Giao Trinh Di Truyen Hoc - Hoang Trong Phan, Truong Thi Bich Phuong, Tran Quoc Dung

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vn
Mc lc
Li ni u
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Hong Trng Phn
M u
I. Khi nim di truyn hc II. Lc s pht trin ca di truyn hc III. i tng v cc lnh vc nghin cu ca di truyn hc IV. Cc phng php nghin cu ca di truyn hc V.Cc nguyn tc nghin cu v phng php hc tp di truyn hc VI. Di truyn hc vi cng ngh sinh hc, tin hc v cc vn x hi 13 13 13 17 18 20 21 24 24 25 25 26 26 26 27 28 28 28 29 30 31 32 33 33 34 39
Chng 1: C s ca Di truyn hc Mendel

Hong Trng Phn
I. Tiu s Mendel - Cha ca di truyn hc II. i tng v phng php th nghim ca Mendel 1. i tng 2. Phng php III. Lai mt tnh v nguyn l phn ly 1. Kt qu th nghim lai mt tnh 2. Gii thch v kim chng nguyn l phn ly 3. Nguyn l phn ly v tnh ph bin ca n IV. Lai hai tnh v nguyn l phn ly c lp 1. Kt qu th nghim lai hai tnh 2. Gii thch v ni dung nguyn l phn ly c lp V. S di truyn Mendel ngi 1. Cc tnh trng ln 2. Cc tnh trng tri VI. L thuyt xc sut trong d on v phn tch di truyn hc 1. Mt s khi nim v tnh cht c bn ca xc sut 2. Mt s nguyn l xc sut c bn VII. Phng php 2 (Chi-square method) trong nh gi ph hp gia cc s liu quan st v k vng
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Chng 2: M rng v p dng ca Di truyn hc Mendel
Hong Trng Phn
I. Cc kiu quan h gia cc gene allele i vi mt tnh trng 1. Cc kiu tri hon ton, khng hon ton v ng tri 2. Tc ng ca gene gy cht (lethals) 3. Hin tng a allele (multiple allelism) II. Tnh a hiu ca gene (pleiotropy) III. Cc kiu tng tc gia cc gene khng allele 1. Tng tc b tr (complementary) 2. Tng tc t ch (epistasis) 3. Tng tc cng gp- s di truyn a gene v cc tnh trng s lng IV. Cc mi quan h kiu gene - kiu hnh 1. Thng bin v mc phn ng 2. thm nhp (penetrance) v biu hin (expressivity)
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Chng 3: C s T bo ca s Sinh sn, Di truyn v Bin d

Hong Trng Phn
I. Sinh sn hu tnh v tnh n nh ca cc b nhim sc th II. Hnh thi hc nhim sc th eukaryote 1. Kch thc nhim sc th 2. Tm ng v cc kiu nhim sc th 3. Cc kiu bng nhim sc th (chromosomal bands) III. Chu k t bo v nguyn phn 1. Chu k t bo (cell cycle) 2. Nguyn phn (mitosis) IV. Gim phn, s pht sinh giao t v th tinh 1. Gim phn (meiosis) 2. S pht sinh giao t (gametogenesis) 3. S th tinh (fertilization) V. Cc bin i ca nhim sc th 1. Cc bin i v cu trc nhim sc th 2. Cc bin i v s lng nhim sc th
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Chng 4: Di truyn hc Nhim sc th
Hong Trng Phn 103
I. Trng phi Morgan vi thuyt di truyn nhim sc th 1. Tm quan trng ca rui gim Drosophila 2. Thuyt di truyn nhim sc th II. S xc nh gii tnh (sex determination) 1. S xc nh gii tnh do kiu gene (GSD) 2. S xc nh gii tnh do mi trng (ESD) III. S di truyn lin kt vi gii tnh (sex-linked inheritance) 1. c im di truyn ca cc gene trn nhim sc th X v Y 2. S bt hot ca nhim sc th X v mt s vn lin quan 3. Cc tnh trng gii hn bi gii tnh v chu nh hng ca gii tnh IV. Lin kt v ti t hp ca cc gene trn mt nhim sc th 1. Khm ph v s trao i cho rui gim 2. Lin kt gene hon ton (gim phn khng c trao i cho) V. Trao i cho v lp bn di truyn 1. Tn s ti t hp 2. Bn di truyn 3. Trao i cho bn si VI. Lp bn gene t cc php lai phn tch ba im 1. Trao i cho kp vi vic xc nh trt t v khong cch cc gene 2. nhiu v h s trng hp (coefficient of coincidence) VII. Lp bn bng phn tch b bn (tetrad analysis)
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Chng 5: Bn cht Ho hc v Ti bn ca Vt cht Di truyn

Hong Trng Phn 147
I. Bng chng vt cht di truyn l cc nucleic acid 1. Cc th nghim bin np vi khun 2. Tnh n nh ca hm lng DNA cc sinh vt bc cao 3. Cc th nghim virus II. Thnh phn ho hc v cu trc ca DNA 1. Cu trc ca mt nucleotide
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2. Cu trc chui polynucleotide 3. Thnh phn ho hc v cu trc ca chui xon kp DNA 4. S lc v cc c tnh ho l ca cc nucleic acid III. Kch thc b gene v tnh phc tp v mt tin ho 1. S lc v b gene ca cc virus, prokaryote v eukaryote 2. Mi quan h gia kch thc b gene v tnh phc tp v tin ho 3. Hm lng DNA v nghch l gi tr C 4. V kch thc DNA cc bo quan IV. T chc phn t ca cc nhim sc th 1. Cu trc cht nhim sc 2. S lc cc thnh phn DNA trong b gene eukaryote V. Ti bn DNA (DNA replication) 1. Cc nguyn tc v c im chung ca ti bn DNA 2. Cc enzyme tham gia ti bn DNA 3. C ch ti bn DNA VI. Ti bn ca cc b gene RNA (RNA genomes) 1. c im ti bn ca cc b gene RNA virus 2. Ti bn ca b gene RNA 3. Phin m ngc
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Chng 6: Gene v Qu trnh Sinh tng hp Protein

Hong Trng Phn 179
I. S pht trin ca khi nim gene 1. Cc quan nim ca Mendel v Morgan v gene 2. Gi thuyt mt gene - mt enzyme ca Beadle v Tatum 3. Quan nim ca Benzer v cc n v cu trc v chc nng di truyn 4. Mi quan h gene - cistron cc prokaryote v eukaryote II. Cu trc v chc nng ca protein 1. Cu trc protein 2. Chc nng protein III. M di truyn 1. Bng chng di truyn hc v m b ba 2. Gii m di truyn
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3. Cc c tnh ca m di truyn 4. Nhng ngoi l so vi m di truyn "ph bin" 5. S linh hot trong vic kt cp anticodon-codon IV. C ch phin m (transcription) v sa i sau phin m 1. Cc RNA v c im chung ca phin m 2. Cc RNA polymerase ca prokaryote v eukaryote 3. Cc promoter cc prokaryote v eukaryote 4. Cc giai on ca qu trnh phin m 5. S sa i sau phin m i vi cc mRNA eukaryote V. Cu trc v chc nng ca cc loi RNA v ribosome 1. RNA thng tin (messenger RNA = mRNA) 2. RNA vn chuyn (transfer RNA = tRNA) 3. RNA ribosome (ribosomal RNA = rRNA) 4. Ribosome VI. C ch dch m (translation) 1. Hot ho amino acid 2. C ch ca qu trnh dch m (tng hp polypeptide)
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Chng 7: S iu ho Biu hin ca Gene

Trng Th Bch Phng 208
I. Cc nguyn l iu ho v mc kim sot phin m II. iu ho biu hin gene prokaryote 1. Cu trc ca operon 2. iu ho dng tnh operon lactose 3. iu ho m tnh operon tryptophan 4. Phin m d (attenuation) III. iu ho biu hin gene eukaryote 1. S bin i DNA 2. Cc promoter 3. Nhng trnh t tng cng phin m (enhancer) 4. Trnh t bt hot gene (gene silencing) 5. Promoter chn lc (alternative promoter) 6. Splicing chn lc
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Chng 8: t bin Gene, Ti t hp v cc Yu t Di truyn Vn ng
I. t bin gene 1. Cc kiu t bin gene 2. Cc tc nhn gy t bin gene 3. C ch phn t ca cc t bin gene II. Sa cha v bo v DNA III. Cc yu t di truyn vn ng (transposable genetic elements) 1. Cc yu t di truyn vn ng prokaryote 2. Cc yu t di truyn vn ng eukaryote
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Chng 9: S Di truyn T bo cht

I. S di truyn t bo cht 1. S di truyn ca cc gene lp th 2. S di truyn ca cc gene ty th 3. Hiu qu dng m ln chiu xon v c II. Lp bn ty th v lp th 1. Lp bn gene ca DNA lp th 2. Lp bn gene ca DNA ty th III. Di truyn hc phn t cc bo quan 1. Cc b gene lp th (cpDNA) 2. Cc b gene ty th (mtDNA)
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Hong Trng Phn 258
Chng 10: i cng v Cng ngh DNA Ti t hp

I. Cc cng c chnh ca k thut to dng DNA ti t hp 1. Cc enzyme gii hn 2. Cc vector thng dng trong k thut di truyn 3. Thit lp phn t DNA ti t hp in vitro II. To dng gene hay DNA ti t hp 1. Nguyn tc chung 2. Quy trnh to dng gene ti t hp 3. Tng hp v to dng cDNA 258 258 260 261 263 263 264 267

III. Cc phng php biu hin cc gene c to dng IV. ng dng ca cng ngh DNA ti t hp 1. Cng ngh DNA ti t hp vi vic nghin cu b gene 2. Cng ngh DNA ti t hp vi y-dc hc 3. K thut di truyn vi cc sinh vt bin i gene
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Chng 11: Di truyn hc Ngi

Trn Quc Dung 278
I. Cc phng php nghin cu di truyn hc ngi 1. Phng php phn tch ph h (genealogy analysis) 2. Phng php nghin cu tr sinh i 3. Phng php di truyn t bo hc ngi 4. Phng php nghin cu qun th 5. Cc k thut sinh hc phn t II. Cc phng php lp bn di truyn ngi 1. Phn tch lin kt (linkage analysis) 2. Cc phng php lp bn vt l (physical mapping) III. Nhim sc th Y v cht nhim sc gii tnh ca ngi 1. Nhim sc th Y ca ngi 2. Cht nhim sc th gii tnh ca ngi IV. S di truyn cc gene tri-ln trn nhim sc th thng v nhim sc th gii tnh 1. S di truyn cc gene tri-ln trn nhim sc th thng 2. S di truyn cc gene tri-ln trn nhim sc th gii tnh V. Di truyn y hc 1. Cc bnh di truyn do ri lon chuyn ho v cc bnh nhim sc th 2. C s di truyn ung th VI. T vn di truyn y hc
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Chng 12: Di truyn hc Qun th

Hong Trng Phn 296
I. Cc khi nim c bn ca di truyn hc qun th 1. Qun th (population) 2. Cc h thng giao phi
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3. Vn gene (gene pool) 4. Tn s kiu gene v tn s allele II. Nguyn l Hardy-Weinberg v trng thi cn bng ca qun th 1. Nguyn l Hardy-Weinberg 2. Nhng ng dng ca nguyn l Hardy-Weinberg III. M rng nguyn l Hardy-Weinberg 1. a allele (multiple alleles) 2. Tn s allele sai bit gia hai gii tnh 3. Cc gene lin kt trn X IV. Ni phi (inbreeding) 1. T th tinh (self-fertilization) 2. H s ni phi (inbreeding coefficient) 3. Tnh ton h s ni phi V. Cc nhn t tc ng ln thnh phn di truyn qun th 1. t bin 2. Bin ng di truyn ngu nhin 3. Dng gene hay s di nhp c 4. Chn lc t nhin
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Li ni u
n nay, di truyn hc ra i ch mi hn mt trm nm song n pht trin vi mt tc ht sc nhanh chng. c bit l, trong vng 50 nm li y k t ngy James Watson v Francis Crick khm ph ra cu trc phn t DNA, 25/4/1953. S hon thnh vic gii m di truyn bi hai nhm nghin cu ca Marshall Nirenberg v Har Gobind Khorana vo thng 6 nm 1966 v s ra i ca K thut Di truyn v Cng ngh DNA ti t hp vo gia thp nin 1970 l hai s kin ni bt nht k t sau khi sinh hc phn t ra i. K , s hon tt ca D n B gene Ngi vo thng 4 nm 2003 c xem l mt trong nhng k cng thm him v i nht ca loi ngi. Ln u tin con ngi c th c c mt cch y ton b trnh t 3.164.700.000 cp base trong b gene ca mnh. Tt c nhng s kin ni bt ny minh chng mt iu rng: S pht trin cng vi nhng thnh tu t c ca di truyn hc trong thi gian qua qu l v cng to ln! gp phn i mi ni dung gio trnh Di truyn hc theo hng cp nht kin thc cng nh phng php dy v hc b mn bc i hc, chng ti tham cu nhiu ti liu khc nhau v n lc bin son gio trnh trn tinh thn y. Chng ti hy vng rng gio trnh ny s p ng c phn no nhu cu ging dy v hc tp ca ging vin v sinh vin, v cng c th s dng nh mt ti liu tham kho b ch cho gio vin Sinh hc cc trng THPT trong bi cnh i mi gio dc hin nay. Ni dung gio trnh gm phn M u cng vi 12 chng bao qut cc kin thc i cng ca mt gio trnh Di truyn hc. Cc chng 14 cp ch yu ni dung thuc Di truyn hc c in, cc chng 5-10 tp trung vo phn Di truyn hc phn t v chng 12 c xem l phn nhp mn ca Di truyn hc qun th, cn chng 11 l s kt hp gia cc kin thc di truyn c in v hin i trn i tng l con ngi. Cui mi chng u c cc phn Cu hi v Bi tp v Ti liu Tham kho bn c tin n tp v tra cu. Gio trnh Di truyn hc c ra i trong khun kh ca D n Gio dc thuc i hc Hu, v vy mt s kin thc nng cao s c cp trong mt gio trnh ring, nh: Di truyn Vi sinh vt v ng dng,v Cng ngh DNA Ti t hp. Bn cnh , mt s thut ng khoa hc c thng nht s dng bng ting Anh gip ngi hc d dng hn trong vic tip cn vi thng tin qua sch bo nc ngoi hoc internet.
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Gio trnh ny do ThS. Hong Trng Phn (ch bin), TS. Trng Th Bch Phng v TS. Trn Quc Dung l nhng ging vin ang cng tc ti Khoa Sinh hc cc trng i hc S phm v i hc Khoa hc thuc i hc Hu bin son, vi s phn cng nh sau: ThS. Hong Trng Phn bin son phn M u v cc chng 1, 2, 3, 4, 5, 6, 10 v 12; TS. Trng Th Bch Phng bin son cc chng 7, 8 v 9; v TS. Trn Quc Dung bin son chng 11. gio trnh ny kp thi ra mt bn c, chng ti xin trn trng cm n D n Gio dc i hc Hu ti tr cho vic bin son v xut bn gio trnh trong khun kh ca D n Gio dc i hc mc B. Chng ti xin by t lng cm n c bit n GS. TS. Phan C Nhn, Trng i hc S phm H Ni, dy cng c bn tho v cho nhiu kin qu bu cng nh khch l chng ti rt nhiu k t khi cng gio trnh bt u c hnh thnh. Do kh nng cn hn ch, chc chn gio trnh cn nhiu thiu st. Chng ti rt mong nhn c s ph bnh v ch bo ca cc ng nghip v bn c gio trnh c hon chnh hn trong ln in sau. Hu, ngy 25 thng 6 nm 2005 Cc tc gi

M u
I. Khi nim di truyn hc
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Theo quan nim ca Bateson (1906), di truyn hc (genetics) l khoa hc nghin cu cc c tnh di truyn v bin d vn c ca mi sinh vt cng vi cc nguyn tc v phng php iu khin cc c tnh . y, tnh di truyn (heredity) c biu hin s ging nhau gia con ci vi cha m; v tnh bin d (variability) biu hin s sai khc gia cha m v con ci cng nh gia cc con ci vi nhau. Cn lu rng, gene l khi nim cn bn ca di truyn hc cho nn ni dung ca khi nim gene khng ngng c pht trin cng vi s pht trin ca di truyn hc.
II. Lc s pht trin ca di truyn hc

S ra i v pht trin ca di truyn hc gn lin vi cng trnh nghin cu ca Gregor Mendel nm 1865. Tuy nhin, trc thi Mendel c bit l t th k XVII c mt s s kin quan trng sau y: (1) S ra i ca knh hin vi s khai bi A.van Leuvenhook (1632-1723); v (2) Sinh hc bt u pht trin mnh vo th k XIX vi s ra i thuyt t bo ca M.Schleiden v T.Schwann (1838,1839) v cc thuyt tin ha ca J.B.Lamarck (1809) v c bit l ca R.C.Darwin (1859). Nhn chung, quan nim ph bin thi by gi vn l s di truyn cc tnh trng tp nhim (inheritance of acquired characters) do Lamarck xut v s di truyn ha hp (blending inheritance), ngha l s pha ln "tinh cha huyt m" con ci. 1. S ra i v pht trin ca di truyn Mendel T u H Lan (Pisum sativum), vi tng v phng php nghin cu c o, nm 1865 Gregor Mendel (Hnh 1) pht hin ra cc quy lut di truyn c s u tin v qua suy ra s tn ti tt yu ca cc n v i truyn c th - nhn t di truyn (genetic factor) - quy nh cc tnh trng c truyn t th h ny sang th h khc m sau ny gi l gene. Tuy nhin, gii khoa hc ng thi khng hiu v do khng th nh gi tm vc v i ca pht minh ny. Hnh 1 G. Mendel Mi n nm 1900, ba nh thc vt hc l Carl Correns (Germany), Hugo de Vries (Netherlands) v Erich von Tschermak (Austria) c lp nhau khm ph li cc quy lut di truyn ca Mendel. V di truyn hc chnh thc ra i t y m ngi sng lp l Mendel.
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Trong nhng nm u th k XX, nh ng dng di truyn Mendel, cc nh chn ging pht hin thm cc hin tng nh: tri khng hon ton, ng tri, gene gy cht, a allele, cc kiu tng tc gene... giai on ny, ngoi thuyt t bin ca H.de Vries nm 1901, cn c hai s kin lin quan n s ra i ca thuyt di truyn nhim sc th v di truyn hc qun th sau ny, l s khi xng "thuyt nhim sc th" bi Walter Sutton v Theodor Bovary nm 1902 v vic thit lp quy lut Hardy-Weinberg nm 1908. Mt s thut ng thng dng cng c xut trong giai an ny, nh: di truyn hc (genetics) bi W.Bateson nm 1906, gene, kiu gene (genotype) v kiu hnh (phenotype) bi W.Johannsen nm 1909. 2. S ra i v pht trin ca thuyt di truyn nhim sc th T 1910, Thomas Hunt Morgan (Hnh 2) cng vi ba cng s l Alfred H.Sturtevant, Calvin Bridges v Herman J. Muller xy dng thnh cng thuyt di truyn nhim sc th (chromosome theory of inheritance) da trn i tng nghin cu l rui gim Drosophila melanogaster. Hc thuyt ny xc nhn rng gene l n v c s ca tnh di truyn nm trn nhim sc th ( trong nhn); trn cc gene sp xp theo ng thng
Hnh 2 T.H.Morgan
to thnh nhm lin kt. Nhng ng gp ng k ca cc mn xut sc ca Morgan l: xy dng bn di truyn (Sturtevant 1913), ch ra c ch xc nh cc kiu hnh gii tnh rui gim (Bridges 1916) v pht trin phng php gy t bin bng tia X (Muller 1927). Vi ng gp to ln Morgan c trao gii Nobel nm 1933 v Muller nm 1946. Nm 1931, Barbara McClintock (Hnh 3) v Harriet Creighton thu c bng chng vt l trc tip v ti t hp ng. Sau , hin tng ny cng c C. Stern quan st Drosophila. Nh vy ti t hp c th c pht hin c v mt vt l ln di truyn ng vt cng nh thc vt. n 1944, McClintock pht hin cc yu t di truyn vn ng (transposable genetic elements), v b c trao gii Nobel nm 1983 v khm ph ny.
Hnh 3 B.McClintock
3. S ra i v pht trin ca di truyn hc phn t S ra i ca di truyn hc phn t (molecular genetics) gn lin vi cc khm ph v DNA (deoxyribonucleic acid) t gia th k XX trn i
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tng nghin cu ch yu l cc vi sinh vt. Tuy nhin, trc Friedrich Miescher (1869) khm ph ra mt hn hp trong nhn t bo gi l nuclein m thnh phn chnh ca n sau ny c bit l DNA.
Hnh 4 Beadle, Tatum, Jacob v Monod (t tri sang)
V mi quan h gia gene v protein, t 1902 Archibald Garrod qua nghin cu bnh alcaptonuria ngi gi rng y l mt tnh trng ln Mendel, c th lin quan ti s sai hng mt enzyme. Bng cc th nghim gy t bin cc gene lin quan n cc con ng sinh ha trn nm mc Neurospora, nm 1941 George Beadle v E.L.Tatum (Hnh 4) xc nhn mi gene kim sot s tng hp mt enzyme c th. Chnh gi thuyt mt gene-mt enzyme (one gene-one enzyme hypothesis) ni ting ny m ng cho s ra i ca di truyn ha-sinh, v hai ng c trao gii Nobel cng vi Joshua Lederberg nm 1958. V sau, gi thuyt ny c chnh xc ha l mt gene xc nh ch mt chui polypeptid cu trc s cp ca cc protein, trong c cc enzyme. Vy bn cht ca gene l g? Nm 1944, Oswald Avery (Hnh 5) v cc cng s l MacLeod v McCarty bng th nghim bin np in vitro chng minh rng DNA l vt cht mang thng tin di truyn. Nm 1949, Erwin Chargaff cng b cc kt qu u tin v thnh phn ha hc ca DNA mt s loi.
Hnh 5 O.T. Avery
Vic nghin cu cu trc phn t DNA c bt u t 1951 vi cc dn liu nhiu x tia X ca Rosalind Franklin v Maurice Wilkins (Hnh 6). Cc s liu ha hc v vt l ny l c s m t James Watson v Francis Crick (Hnh 7) xy dng thnh cng m hnh cu trc phn t DNA nm 1953, cn gi l chui
Hnh 6 R.Franklin (tri) v M.Wilkins
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xon kp (double helix). Pht minh v i ny m ra k nguyn mi cho s pht trin ca di truyn hc v sinh hc ni chung. Vi pht minh , Watson v Crick cng vi Wilkins c trao gii Nobel nm 1962 . Bi bo nhan "Mt cu trc cho Deoxyribose Nucleic Acid" ca Watson v Crick ng trn tp ch Nature ngy 25/4/1953 c nh gi
Hnh 7 J.D.Watson (tri) v F.H.C.Crick
l mt bi bo khng bnh thng. Ch vn vn c 128 dng nhng ng sau bi bo l c mt bc tin lch s v i ca di truyn hc m mi dng l mt cu chuyn. Tht vy, sau cu trc chui xon kp l hng lot cc khm ph mi. Nm 1958 Matthew Meselson v Franklin Stahl chng minh s ti bn bn bo ton ca DNA; v nm 1961 Seymour Benzer hon tt cng trnh nghin cu cu trc tinh vi ca gene; Francois Jacob v Jacques Monod (Hnh 4) tm ra c ch iu ha sinh tng hp protein (gii Nobel 1965 vi Andre Lwoff); S.Brenner, Jacob v Meselson khm ph ra RNA thng tin; S.Brenner v F.Crick chng minh m ditruyn l m b ba; cng trnh gii m
Hnh 8 H.G.Khorana (tri) v M.Nirenberg
di truyn ny c hon thnh vo thng 6 nm 1966 bi hai nhm nghin cu ca Marshall Nirenberg v Har Gobind Khorana (gii Nobel nm 1968; Hnh 8). 4. S ra i v pht trin ca cng ngh DNA ti t hp C th ni, nn tng ca cng ngh DNA ti t hp (recombinant DNA technology) c thnh lp t 1972 khi Paul Berg (Hnh 10) to ra phn t DNA ti t hp u tin trong ng nghim (recombinant DNA in vitro). Mt nm sau Herbert Boyer v Stanley Cohen (Hnh 10) ln u tin s dng plasmid to dng DNA. Lnh vc ng dng mi ny ca sinh hc phn t to ra mt cuc cch mng mi trong sinh hc. ng gp ng k trong lnh vc ny l khm ph v cc enzyme gii hn (restriction enzyme) t 1961-1969 ca Werner Arber, Daniel Nathans v
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Hamilton Smith (gii Nobel 1978; Hnh 10); xut cc phng php xc nh trnh t base trong cc nucleic acid nm 1977 bi P.Berg, W.Gilbert
Hnh 9 Cc nh khoa hc ot gii Nobel y hc lin quan k thut gene. T tri sang: D.Nathans, H.Smith, W.Arber, P.Sharp v R.Robert.
Hnh 10 Cc nh khoa hc ot gii Nobel ha hc lin quan k thut gene. T tri sang: H.Boyer, S.Cohen, P.Berg, W.Gilbert, F.Sanger v K.Mullis.
v Frederick Sanger (gii Nobel ha hc 1980; Hnh 10); s khm ph ra cc gene phn on (split gene) nm 1977 bi Phillip Sharp v Richard Robert (gii Nobel 1993; Hnh 9); s pht minh ra phng php PCR (polymerase chain reaction) ca Kary B.Mullis nm 1985 (Hnh 10) v phng php gy t bin nh hng (site-directed mutagenesis) ca Michael Smith t 1978-1982 (gii Nobel ha hc 1993)... Cng vi nhng thnh tu ng dng ly k trong sn xut v i sng x hi, nh vic sn xut cc ch phm y-sinh hc bng cng ngh DNA ti t hp, s dng liu php gene (gene therapy) trong iu tr bnh di truyn, to cc ging sinh vt mi bng con ng bin i gene (genetically modified organisms = GMOs), d n b gene ngi (Human Genome Project = HGP)... gy ra khng t hoi nghi, tranh ci xung quanh cc vn v o l sinh hc (bioethics) v an ton sinh hc (biosafety).
III. i tng v cc lnh vc nghin cu ca di truyn hc

Trong giai on u, i tng ca di truyn hc l cc thc vt, ng vt, ngi v cc vi sinh vt. T dn ti s hnh thnh cc lnh vc nghin cu tng ng l di truyn hc thc vt, ng vt, ngi v di truyn hc vi sinh vt, trong di truyn hc t bo l c s. Giai on
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ny ko di t thi Mendel cho n thp nin 1940, vi c trng l nghin cu quy lut di truyn cc tnh trng qua cc th h. V vy n thng c gi l giai on di truyn hc Mendel hay di truyn hc c in (Mendelian or classical genetics). T thp nin 1950 n nay, vi s ra i ca di truyn hc phn t (molecular genetics), i tng nghin cu l t chc, cu trc, chc nng v c ch hot ng ca cc b gene (genomes), cc gene v cc sn phm ca chng mc phn t. c bit vi s ra i ca cc k thut to dng gene (gene-cloning techniques) t thp nin 1970, vic nghin cu c bn cng nh ng dng tr nn ht sc thun li. S phn ha thnh cc chuyn ngnh lc ny l v cng phong ph v a dng, nh: di truyn hc bnh (genetics of disease), di truyn hc ung th (genetics of cancer), di truyn hc pht trin (developmental genetics), cng ngh sinh hc (biotechnology)...Gn y cn xut hin mt s lnh vc nghin cu mi nh genomics, DNA chip technology, DNA microarray technology... Mt hng khc ca di truyn hc chuyn nghin cu cu trc di truyn ca cc qun th v s bin i di truyn bn trong qun th v gia cc qun th. l nhnh di truyn hc qun th (population genetics), m nguyn l c s ca n c G.Hardy (England) v W.Weinberg (Germany) c lp a ra nm 1908. Tuy nhin, lnh vc nghin cu ny ch thc s bt u t thp nin 1930 vi cc cng trnh ca R.A.Fisher, J.B.S.Haldane v Sewall Wright. Ngy nay cc nh di truyn hc khng cn thin v nghin cu s bin i mc kiu hnh m tp trung vo s bin i phn t trong mt qun th nhm tm hiu ngha tin ha ca cc bin i . V nh th di truyn hc qun th tr thnh nn tng cho cc thuyt tin ha hin i. Qua phn tch trn cho thy ba nhnh chnh ca di truyn hc, l: di truyn hc Mendel, di truyn hc phn t v di truyn hc qun th.
IV. Cc phng php nghin cu ca di truyn hc

Vic nghin cu di truyn hc c tin hnh bi nhiu phng php khc nhau. Bn cnh cc phng php kinh in c th cn c s pht trin v tch hp ca cc phng php t ton hc, tin hc, vt l v ha hc c bit l trong lnh vc sinh hc phn t. 1. Cc phng php kinh in c th ca di truyn hc Trc ht l phng php t th phn dng chn cc dng thun lm b m trong cc php lai, cc phng php lai mt hoc ng thi nhiu tnh trng gia cc b m do Mendel xut. Trong bao gm c cc hnh thc lai thun nghch v lai phn tch (testcross) nhm rt
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ra quy lut, kim tra kiu gene hoc dng thit lp bn di truyn. i vi nghin cu di truyn ngi v mt s vt nui giao phi cn huyt (consanguineous), c trng l phng php phn tch ph h (pedigree analysis) nhm xc nh c im di truyn tri-ln ca mt tnh trng hoc bnh tt; nghin cu tr sinh i cng trng v khc trng nhm xc nh h s di truyn (heritability) ca tnh trng; phng php gy t bin (mutagenesis) kt hp vi lai hu tnh dng trong nghin cu v chn to ging, c bit l thc vt... 2. Phng php ton hc Trong nghin cu khoa hc ni chung v di truyn hc ni ring, vic p dng cc cng c ton thng k v l thuyt xc sut phn tch nh lng v l gii cc kt qu nghin cu l rt thit yu. Chnh iu lm cho di truyn hc tr thnh mt khoa hc chnh xc v mang tnh d bo. iu ny th hin r trong cc cng trnh nghin cu ca Mendel, Morgan cng nh trong cc nghin cu di truyn s lng v di truyn qun th. 3. Cc phng php vt l v ha hc Trong nghin cu di truyn, c bit l di truyn phn t v t bo i hi phi s dng cc k thut v phng php ca vt l v ha hc. Chng hn, trong nghin cu hnh thi nhim sc th khng th thiu cc loi knh hin vi quang hc v in t, cc k thut nhum bng (banding techniques); nghin cu thnh phn ha hc v cu trc DNA i hi cc phng php sc k v nhiu x tia X... 4. Cc phng php t bo hc Trong nghin cu di truyn t bo c rt nhiu phng php c s dng quan st hnh thi nhim sc th v thit lp kiu nhn (karyotype) ca cc loi cng nh chn on cc bnh tt lin quan n s bin i s lng v cu trc nhim sc th. Bn cnh cc k thut nui cy m v chun b nhim sc th l cc k thut nhum bng khc nhau, k thut lai hunh quang ti ch (fluorescence in situ hybridization = FISH), k thut min dch t bo hc... (xem Verma v Babu 1995). 5. Cc phng php ca sinh hc phn t - k thut di truyn S tin b nhanh chng gn y ca sinh hc phn t v cng ngh sinh hc l nh s pht trin mnh m ca cc k thut ti t hp DNA, lai phn t, s dng cc mu d v phng php nh du khc nhau, phng php PCR, cc phng php xc nh trnh t nucleic acid cng nh cc phng php bin i vt liu di truyn mi. Vi cc cng c k thut mi ny cho php i su nghin cu t
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chc ca cc gene v b gene, cc c ch iu ha v bin i di truyn mc phn t cng nh cc thnh tu ng dng mi trong y-sinh hc, nng nghip v cc lnh vc khc ca i sng-x hi. V. Cc nguyn tc nghin cu v phng php hc tp di truyn hc 1. Cc nguyn tc nghin cu ca di truyn hc Trong nghin cu di truyn hc v sinh hc ni chung c cc nguyn tc chung cn tun th nh l phng php lun, sau y: (1) Ly t bo lm n v nghin cu; (2) Thng tin di truyn cha trong b gene t bo chi phi mi biu hin sng ca n m cc gene l n v di truyn c s; (3) S hot ng ca cc gene trong qa trnh pht trin c th l c trng cho tng gene trong tng giai on c th; (4) Cc qu trnh trong cc h thng sng phi c iu ha v kim sot m bo cho s tn ti ca n l lin tc, trong ph bin l s t iu chnh bng cc c ch phn hi thng tin (feed-back mechanism); (5) S thng nht gia cu trc v chc nng biu hin tt c cc mc t chc khc nhau ca s sng; (6) Tt c cc t chc v qu trnh sng u tun theo cc quy lut vt l v ha hc; (7) S sng trn tri t tri qua qu trnh tin ha khong 3,5 t nm qua, v vy khi so snh, nhng nt tng ng gia chng cho thy tnh thng nht v mt ngun gc v nhng nt d bit cho thy tnh pht trin, s phn ha a dng tt yu ca chng. 2. Phng php hc tp mn di truyn hc Cng nh bt k mn hc no khc, vic hc tp mn di truyn i hi phi nm vng lch s mn hc, i tng, nhim v, phng php nghin cu v h thng kin thc cn bn ca n. Bn cnh cc nguyn tc ni trn vn rt cn cho t duy trong hc tp, di y nu mt s im chnh lin quan phng php hc tp c th ca b mn. (1) Nm vng cc kin thc lin mn (nh t bo hc, ha sinh hc, vi sinh hc, hc thuyt tin ha...) v lin ngnh (nh cc khi nim v nguyn l c bn ca ton thng k-xc sut, vt l v ha hu c). (2) Nm vng h thng khi nim c bn cng nh cc thut ng mi khng ngng ny sinh. Trong gene l khi nim cn bn c ni hm khng ngng pht trin, c bit l trong ba thp nin li y. (3) Hiu r bn cht ca cc nguyn l di truyn trong tng ch cng nh mi lin quan gia chng c th gii thch v vn dng trong gii quyt cc bi ton hoc tnh hung ca i sng v thc tin sn xut. (4) nm kin thc v pht trin cc k nng t duy mt cch vng chc i hi phi bit vn dng kin thc vo gii bi tp cng nh cc k nng thc hnh th nghim.
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(5) Di truyn hc l mt khoa hc thc nghim, nn thng tin thu c l nh cc quan st t th gii t nhin, v phng php khoa hc chnh l cng c hiu bit cc quan st . Ni n phng php nghin cu khoa hc l ni n cc bc tin hnh theo mt trnh t t chc cng vic cht ch sau y: Quan st Gi thuyt D on Thc nghim ( kim tra gi thuyt t ra) xut gi thuyt mi. Cng cn lu rng, c tnh ca khoa hc l hoi nghi, i hi phi c bng chng xc thc, l kt hp gia logic v tr tng tng, l gii thch v d on, khng c s c on; ngi nghin cu hay nh khoa hc phi nhn bit sng t, trung thc, v t, v ni chung l chu s chi phi ca cc nguyn tc o c c tha nhn rng ri. (6) Trong khi hc gio trnh, bn nn lm t nht mt tiu lun v mt vn cp nht mnh yu thch. iu rt l th v b ch. Bi cng vic ny i hi s say m tm ti cc thng tin mi, c bit l trn mng vit mt bi tng lun c tnh khoa hc v trnh by trong mt seminar. (7) Bi di truyn hc l mt ngnh khoa hc non tr nhng pht trin vi tc cc nhanh, nn khi lng kin thc mi tch ly c l v cng phong ph v a dng. c th cp nht thng tin v mn hc i hi phi tng cng kh nng s dng ting Anh v internet. iu quan trng l phi to cho mnh mt hoi bo hc tp, mt kh nng v phng php t hc v thnh lp cho c mt th mc tra cu. Trong ng k l cc trang web (c gii thiu trong tng chng), hoc c th s dng ngay cc t kha (key words) c cho tng ch tm kim vi cng c c th ni mnh nht hin nay l Google. VI. Di truyn hc vi cng ngh sinh hc, tin hc v cc vn x hi Nh cp, s pht trin ht sc nhanh chng ca di truyn hc trong vi thp nin qua, c bit l s tin b ca cng ngh sinh hc (biotechnology) ni chung c nhng tc ng mnh m ln nhiu ngnh khoa hc v trn mi mt ca i sng, kinh t, chnh tr v x hi phm vi ton cu. Di truyn hc c hnh dung v tr trung tm v giao thoa vi sinh hc, ha sinh hc, k ngh, y-dc, nng nghip, sinh thi hc, kinh t hc, lut, x hi hc v trit hc. Gio s danh d mn ha hc i hc Havard, F.H.Westheimer, bnh lun v sinh hc phn t nh sau:"Cuc cch mng tr tu v i nht ca 40 nm qua xy ra trong sinh hc. Liu c th tm ra mt ngi no c hc ngy nay m khng hiu bit cht g v sinh hc phn t?" (dn theo Weaver v Hedrick 1997, tr.15). Cc thnh tu t c nh ng dng di truyn hc trong nng nghip
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l v cng to ln, gp phn to nn cuc "cch mng xanh ln th hai" vi s ra i ca hng lot cc ging vt nui-cy trng c u th lai vt tri, cc sinh vt bin i gene (GMOs) mang nhng c tnh hon ton mi l. Trong y hc, l s ra i ca hng lot cc dc phm c sn xut bng k thut di truyn dng cho iu tr bnh v ci bin tr thng minh ca con ngi; l cc phng php chn on v iu tr bnh mc phn t...S thnh cng ca d n b gene ngi (HGP) vo thng 4 nm 2003 cho php chng ta ln u tin c c ton b trnh t khong 3,2 t cp nucleotide trong b gene con ngi (Homo sapiens). HGP l mt trong nhng k cng thm him v i nht trong lch s nhn loi (NHGRI 2005). Theo c tnh mi nht c cng b ngy 21/10/2004 trn tp ch Nature, b gene chng ta cha s lng gene m ha protein thp mt cch ng kinh ngc, khong 20.000 n 25.000 ch khng phi l 50.000 n 140.000 gene nh d on ban u hoc 35.000 theo d on trong vi ba nm li y (NHGRI 2005). Chnh s kt hp tin hc v my tnh trong nghin cu sinh hc phn t dn ti s ra i mt ngnh mi l sinh-tin hc (bioinformatics) cho php thu thp, t chc v phn tch s lng ln cc s liu sinh hc nh s dng mng my tnh v cc ngun d liu (databases). V mt s lnh vc nghin cu mi khc cng ra i nh: genomics - phn tch ton b genome ca mt sinh vt c chn, DNA microchip technology - xc nh cc t bin trong cc gene, DNA microarray technology - nghin cu cch thc mt s lng ln cc gene tng tc ln nhau v c ch mng li iu ha ca t bo kim sot ng thi s lng cc k ln cc gene... Nhng thch thc cho tng lai ca nghin cu khoa hc v cc b gene (genomics) i vi sinh hc, vn sc khe v x hi cng c t ra (Collins v cs 2003). S hon tt ca HGP t n khng c ngha l xong m ng hn l im khi u cho cng cuc nghin cu thm ch cn hng th hn. Cc nh nghin cu hin gi ang c gng lm sng t mt s qu trnh phc tp nht ca sinh hc, l: mt a b pht trin t mt t bo n l bng cch no, cc gene phi hp chc nng ca cc m v c quan nh th no, s tin nh bnh tt xy ra nh th no v b no ngi lm vic ra sao (NHGRI 2005). Cng vi nhng thnh tu ng dng ly k trong sn xut v i sng x hi ni trn, nhiu vn mi c t ra cho cc ngnh gio dc, lut, trit hc, x hi hc v gy khng t hoi nghi, tranh ci xung quanh cc vn v o l sinh hc, an ton sinh hc v mi sinh.

Ti liu Tham kho
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Ting Vit H Hunh Thy Dng. 1997. Sinh hc Phn t. NXB Gio Dc. Phm Thnh H. 2000. Di truyn hc. Ti bn ln II, NXB Gio Dc. Phan C Nhn (ch bin), Nguyn Minh Cng, ng Hu Lanh. 1999. Di truyn hc. NXB Gio Dc. Ting Anh Collins FS, Green ED, Guttmacher AE, Guyer MS. 2003. A vision for the future of genomics research. Nature, Vol. 422, No. 6934, p.835-847. McKusick V. 1998. Mendelian Inheritance in Man. 12th ed., Johns Hopkins University Press, Baltimore. National Human Genome Research Institute (NHGRI)/ National Institute of Health (NIH). 2005: http://www.genome.gov/ http://www.ncbi.nlm.nih.gov/ Online Mendelian Inheritance in Man (OMIMTM): http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=OMIM Tamarin RH. 1999. Principles of Genetics. 6th ed, McGraw-Hill, Inc, NY. Verma RS and Babu A. 1995. Human chromosome : principles and techniques. 2nd ed, international edition, McGraw-Hill, Inc., New York. Watson JD and Crick FHC. 1953. A structure for Deoxyribose Nucleic Acid. April 25,1953, Nature, Vol.171, page 737. Weaver RF, Hedrick PW. 1997. Genetics. 3rd ed, McGraw-Hill Companies, Inc. Wm.C.Brown Publishers, Dubuque, IA. Zinnen T. 2005. JD Watson and FHC Crick - A structure for Deoxyribose Nucleic Acid - Nature, 25 April 1953. (http://www.accessexcellence.org)

Chng 1
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C s ca Di truyn hc Mendel
Cho n u th k XX, mi ngi cn cha hiu c c ch ca s di truyn, mc d vn bit rng con ci sinh ra thng ging b m. Quan nim ph bin cho n gia th k XIX c gi gn trong ci gi l thuyt di truyn ha hp (theory of blending inheritance) nhm gii thch s kin con ci mang cc c im ca c hai b m. Tuy nhin, n nm 1866 Gregor Mendel a ra thuyt di truyn gin on (theory of particulate inheritance), vi gi rng: n v di truyn c th kim sot mt tnh trng c truyn t th h ny sang th h khc tn ti di dng ht, ngy nay ta gi l gene. Cc khm ph quan trng ca Mendel t nn mng cho s ra i ca di truyn hc sau ny. Nh Thomas Hunt Morgan nhn nh: "Trong mi nm nghin cu cy u H Lan trong ngi vn ca tu vin, G. Mendel lm nn s khm ph v i nht trong sinh hc t c trong nm trm nm qua".
I. Tiu s Mendel - Cha ca Di truyn hc

Gregor Mendel sinh nm 1822, ln ln trang tri ca cha mnh ti mt tnh ca Austria (o). V gia nh ngho nn ng phi vo tu vin tip tc vic hc ca mnh. Trong khong thi gian ny, Mendel nghin cu vt l v ton l nhng mn hc gip ng nhiu trong cc th nghim di truyn sau ny. ng c gi ti i hc Vienna thi ly bng gio vin chnh thc, nhng khng v quay v tu vin "dy hc" trong nhiu nm.
Hnh 1.1 Gregor Mendel trong ngi vn ca tu vin Brno.
Khi cn trang tri ca cha mnh, Mendel quan tm ti cc cy ci v con vt, v thng gi li nhng ci hoa, con ong v chut. Sau ny tu vin Brno ng tp trung vo cc cy u H Lan (Pisum sativum). Mendel xc nh c cc nguyn l di truyn c s t cc th nghim chn ging thc vt. Cc kt qu nghin cu ny c Mendel trnh by nm 1865 trc Hi Nghin cu Khoa hc T nhin Brno v cng b nm 1866 Germany trong mt bi bo nhan l Cc th nghim lai thc vt (Experiments on Plant Hybrids). Bi bo ny nhanh chng c mt nhiu th vin, nhng nhng ngi cng thi ng khng
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hiu c cc pht hin ca ng, c l mt phn l do ng s dng ton hc l gii cc kt qu ca mnh. Ngoi ra, hu ht cc nh nghin cu ng thi do tin hnh nhiu tnh trng ng thi dn ti cc kt qu ri bi nn khng th nhn ra c cc nguyn l di truyn c s. Mendel tr thnh tu vin trng t nm 1868 v khng cng b thm mt kt qu no v di truyn na k t sau kit tc nm 1866. Mendel qua i nm 1884 trc khi cng trnh ca ng c gii khoa hc thu hiu. Mi n nm 1900, cng trnh ca ng mi c ba nh thc vt hc c lp nhau khm ph li, l Carl Correns ca Germany, Hugo de Vries ca Netherlands v Erich von Tschermak ca Austria. y l mc khi u cho cc nghin cu di truyn hc hin i. Ngy nay, phng php th nghim ca Mendel c xem l th d kinh in v s nghin cu khoa hc c lp k hoch cn thn v bi bo ca ng l s minh ha tuyt vi ca mt thin ti khoa hc.
II. i tng v phng php th nghim ca Mendel

1. i tng
(a) (b) Hnh 1.2 (a) By tnh trng tng phn u H Lan c Mendel nghin cu; dng tri nm bn tri ca mi trng hp. (b) Cu to hoa u, phng php th phn cho v cy u H Lan.
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Mendel chn u H Lan (Pisum sativum) lm i tng nghin cu v chng c hai c im c bn l sai khc nhau v nhiu tnh trng tng phn d quan st (hnh 1.2a) v sinh sn bng li t th phn. Ngoi ra, u c hoa kh ln nn thao tc d dng (hnh 1.2b); c kh nng cho s lng i con nhiu; v nhiu ging u lc by gi c gi tr kinh t cao. 2. Phng php Tnh cht c o ca phng php nghin cu Mendel th hin ch: (1) Chn cc dng thun (pure lines) khc nhau bng cch cho t th phn lin tip nhiu th h dng lm dng b m trong cc php lai; (2) Theo di trc tin kt qu di truyn ring bit ca tng tnh trng qua vi th h, trong th h cy lai th nht hay F1 sinh ra do giao phn gia hai dng b m thun chng khc nhau, cn th h cy lai th hai hay F2 sinh ra t s t th phn ca cc cy lai F1, ri sau mi tin hnh nghin cu s di truyn ng thi ca hai hoc nhiu tnh trng; (3) Khi qut v l gii cc kt qu th nghim thu c bng ton thng k v xc sut; v (4) Kim tra li mt cch cn thn cc gi thuyt khoa hc bng cc php lai thun nghch (reciprocal matings) v lai phn tch (testcross).
III. Lai mt tnh v nguyn l phn ly

1. Kt qu th nghim lai mt tnh (monohybrid cross) Mendel tin hnh by php lai mt tnh khc nhau v cc kt qu thu c c trnh by Bng 1.1.
Bng 1.1 Cc kt qu lai mt tnh ca Mendel F2 TT Kiu hnh P F1 1 Ht trn nhn Trn 5474 trn : 1850 nhn 2 Ht vng xanh Vng 6022 vng : 2001 xanh 3 Hoa ta trng ta 705 ta : 224 trng 4 Qu phng tp Phng 882 phng : 299 tp 5 Qu xanh vng Xanh 428 xanh : 152 vng 6 Hoa dc thn nh Dc thn 651 dc thn : 207 nh 7 Thn cao thp Cao 787 cao : 277 thp T l F2 2,96:1 3,01:1 3,15:1 2,95:1 2,82:1 3,14:1 2,84:1
T tt c cc php lai trn cho thy: Khi b m th h xut pht (P) thun chng khc nhau v mt cp tnh trng tng phn, th th h F1 tt c con lai u biu hin ch mt tnh trng ca b hoc m, tnh trng c gi l tnh trng tri (dominant) v tnh trng kia khng quan st c gi l tnh trng ln (recessive). Sau cho cc con lai F1 t th phn th th h F2 ng thu c c hai kiu hnh (phenotype) ca b m ban u vi t l xp x 3/4 tri v 1/4 ln. Ngoi ra, Mendel cng cho cc cy F2 t th phn ring r v theo di s phn ly th h F3. Kt qu cho thy 1/4 cy ca F2 sinh ra kiu hnh
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ln tt c u l cc cy ln thun chng; iu c ngha l tt c con ci ca chng l ln. Tuy nhin, trong s 3/4 biu hin kiu hnh tri th mt s l tri thun chng, cn s khc th ging nh cc c th F1 ch chng cho i con gm c tri v ln. Nhn chung, c ba kiu c th F2 l: 1/4 tri thun chng, 1/2 tri khng thun chng (cho i con vi t l 3 tri :1 ln) v 1/4 ln thun chng. 2. Gii thch v kim chng nguyn l phn ly T cc kt qu th nghim Mendel kt lun rng, thng qua cc giao t b m truyn cho con ci cc nhn t di truyn (genetic factor) m ngy nay ta gi l gene. Mendel cn gi rng cc nhn t ny tn ti di vi dng bin i (nay gi l cc allele) xc nh cc kiu hnh khc nhau ca cng mt tnh trng. ng gi nh rng mi c th c hai allele ca mi gene, mt ci nhn t giao t ca b v mt ci t giao t ca m. Mc d iu i vi chng ta by gi c v n gin, nhng thi i Mendel khng c ai hiu c n. By gi ta hy xt th nghim 2. Trc tin, quy c cc gene xc nh cc tnh trng tri v ln bng cc ch ci vit hoa v vit thng, chng hn: A - ht vng, v a - ht xanh. Nh vy c ba kiu gene (genotype), trong hai kiu ng hp t (homozygote) - c hai allele ging nhau: AA v aa, tc dng thun chng v mt kiu d hp t (heterozygote) cha hai alele khc nhau: Aa, tc dng lai. V allele vng l tri hn allele xanh, nn c th d hp t Aa c cng kiu hnh vi th ng hp tri AA. P Ht vng (AA) Ht xanh (aa) Giao t P A a F1 Aa (vng) Giao t F1 ( A : a)ci ( A : a)c Khung Punnett
A A a AA Aa a Aa aa
F2
T l kiu gene 1 AA : 2 Aa : 1 aa T l kiu hnh 3 vng (A-) : 1 xanh (aa)
Hnh 1.3 S biu din kt qu lai mt tnh ca Mendel.
Trong gim phn, mi b m thun chng ht vng (AA) v ht xanh (aa) ch cho mt loi giao t mang allele tng ng l A v a. Do kt qu ca s th tinh ch to ra mt kiu d hp t Aa, ngha l tt c con lai
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F1 u c kiu hnh tri ht vng. Khi bc vo gim phn, cc c th F1 d hp t (Aa) s cho hai loi giao t (A v a) vi t l tng ng. cng l thc cht ca nguyn l phn ly (principle of segregation), hay quy lut th nht ca Mendel (Mendel's first law). Kt qu ca s t th tinh ngu nhin gia cc loi giao t c v ci ca F1 ny s cho ra bn kiu t hp F2, vi t l phn ly theo kiu gene l 1AA: 2Aa: 1aa v t l kiu hnh tng ng l 3 vng (A-): 1 xanh (aa). Lu rng thng thng ngi ta s dng khung Punnett (Punnett square) do nh di truyn hc ngi Anh R.C.Punnett a ra nm 1905 xc nh kt qu di truyn ca cc php lai. S biu din kt qu lai mt tnh c nu hnh 1.3. khng nh nguyn l phn ly, Mendel tin hnh hai th nghim: Mt l, cho cc c th d hp t F1 t th phn nh ni trn; v hai l, cho F1 lai ngc tr li vi b hoc m c kiu hnh ln. Php lai vi mt c th ln nh th c gi l lai phn tch (testcross), thay v gi l lai ngc (backcross) bi v n cho php kim tra kiu gene ca mt c th tri l d hp hay ng hp. C s di truyn ca kiu lai ny nh sau: B m Aa (ht vng) aa (ht xanh) Giao t 50% A : 50% a 100% a i con 50% Aa (vng) : 50% aa (xanh) 3. Nguyn l phn ly v tnh ph bin ca n Sau khi cc nguyn l di truyn ca Mendel c khm ph li nm 1900, tnh ph bin ca cc nguyn l Mendel ni chung v nguyn l phn ly ni ring c cc nh nghin cu khng nh bng cch lp li cc php lai ca ng (chng hn gia u ht vng v ht xanh; bng 1.2) cng nh tin hnh cc php lai tng t cc ng vt v thc vt khc.
Bng 1.2 Cc kt qu lai lp li u H Lan Nh nghin cu Mendel (1866) Correns (1900) Tschermak (1900) Bateson (1905) Darbishire (1909)
Tnh ton b
Vng 6.022 1.394 3.580 11.902 109.060

131.958
Nhn 2.001 453 1.190 3.903 36.186

43.733
T l F2 3,01:1 3,08:1 3,01:1 3,05:1 3,01:1

3,02:1
Ni dung chnh ca nguyn l hay quy lut phn ly c th tm lc nh sau: Cc allele l nhng dng khc nhau ca cng mt gene; trong cc th d hp t chng phn ly v cc giao t vi t l tng ng.
IV. Lai hai tnh v nguyn l phn ly c lp

1. Kt qu th nghim lai hai tnh (dihybrid cross)
29
xc nh s di truyn ng thi ca nhiu cp tnh trng, Mendel tin hnh nhiu th nghim khc nhau. Bng 1.3 gii thiu kt qu lai hai tnh gia cc ging u thun chng ht vng-trn v ht xanh-nhn.
Bng 1.3 Cc kt qu lai hai tnh ca Mendel Th h Ptc F1 F2 Kiu hnh ht Vng-trn xanh-nhn Vng-trn Vng-trn Vng-nhn S lng 315 101 108 32
556
Xanh-trn Xanh-nhn
Tng =
T l F2 (quan st) 9,84 3,16 3,38 1,0
T l F2 (k vng) 9 3 3 1
Vi php lai ny, tt c con lai F1 u c kiu hnh tri kp l ht vng v trn. Khi cho F1 t th phn, F2 xut hin 4 kiu hnh l vng-trn, vng-nhn, xanh-trn v xanh-nhn vi t l xp x 9:3:3:1. 2. Gii thch v ni dung nguyn l phn ly c lp Nu xt t l phn ly ca tng tnh trng F2, ta c: 315 + 101 = 416 vng v 108 + 32 = 140 xanh, xp x 3 vng : 1 xanh. Tng t, v hnh dng ht, ta c 315 + 108 = 423 trn v 101 + 32 = 133 nhn, xp x 3 trn : 1 nhn. iu chng t mi tnh trng u tun theo quy lut phn ly 3 tri :1 ln. Bng cch p dng quy tc nhn xc sut ca cc bin c c lp (xem mc VI), ta d dng chng minh c rng s phn ly ca hai t l ny l hon ton c lp nhau nh d on ban u. Tht vy, (3 vng :1 xanh)(3 trn :1nhn) = 9 vng-trn : 3 vng-nhn : 3 xanh-trn : 1 xanh-nhn. Cn lu l t l 9:3:3:1 ny cng c Mendel tm thy trong khi lp li th nghim vi cc tnh trng khc. T ng mi xy dng nn nguyn l phn ly c lp (principle of independent assortment), cn gi l quy lut th hai ca Mendel (Mendel's second law). Ni dung ca nguyn l ny pht biu rng: Cc allele ca cc gene khc nhau th phn ly mt cch c lp vi nhau trong qu trnh hnh thnh giao t (kt qu l to ra t l 9:3:3:1 th h F2 t mt php lai hai tnh). minh ha cho nhng iu trnh by trn y, ta quy c: A - vng, a - xanh, B - trn, b - nhn. Lu : kim tra li gi thuyt phn ly c lp, Mendel tin hnh lai phn tch gia cc cy vng-trn F1 (AaBb) vi cy xanh-nhn (aabb). Kt qu thu c gm 55 vng-trn : 49 vng-nhn : 51 xanh-trn : 53
30
xanh-nhn, tng ng vi t l 1:1:1:1 = (1:1)(1:1). iu chng t cc cy F1 cho bn loi giao t vi t l ngang nhau, ngha l cha hai cp gene d hp phn ly c lp. vng, trn xanh, nhn Ptc Kiu hnh Kiu gene AABB aabb Giao t AB ab F1 Kiu gene AaBb Kiu hnh vng, trn Giao t AB : Ab : aB : ab Khung Punnett
AB AB Ab aB ab AABB AABb AaBB AaBb Ab AABb AAbb AaBb Aabb aB AaBB AaBB aaBB aaBb ab AaBb Aabb aaBb aabb
F2
T l kiu gene
1/16 AABB + 2/16 AaBB + 2/16 AABb + 4/16 AaBb 1/16 AAbb + 2/16 Aabb 1/16 aaBB + 2/16 aaBb 1/16 aabb
T l kiu hnh
= 9/16 vng, trn = 3/16 vng, nhn = 3/16 xanh, trn = 1/16 xanh, nhn
Hnh 1.4 C s di truyn hc ca nguyn l phn ly c lp.
V. S di truyn Mendel ngi

Cng nh u H Lan, rui gim hay mo, ngi c rt nhiu tnh trng di truyn theo cc quy lut Mendel. Chng hn, theo thng k ca Victor A.McKusick nm 1994, c 6.678 tnh trng v cc bnh n gene. Cho n ngy 8/2/2005, cc s liu v s lng gene v kiu hnh c nu bng 1.4 (OMIM 2005). Ni chung, vic xc nh phng thc di truyn ngi l tng i kh khn, v mi gia nh c t con, thng khng qu 10 ngi. khc phc iu ngi ta s dng phng php phn tch ph h (pedigree analysis). Di y nu mt s tnh trng tri v ln ngi m khng phn tch c im ca cc kiu di truyn (xem chng 11).

Bng 1.4 S lng cc mc NST thng 9517 360 1512 1326 2150 14.865 Lin kt X 423 38 137 134 153 885 Lin kt Y 48 0 2 4 2 56 DNA ty th 37 0 27 0 0 64
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Gene c trnh t bit Gene c trnh t v kiu hnh r M t kiu hnh, c s ph.t r Kiu hnh hay locus Mendel, c s phn t cha bit Cc kiu hnh chnh yu khc c c s Mendel cn kh nghi Tng
Tng 10.025 398 1.678 1.464 2.305 15.870
1. Cc tnh trng ln (recessive traits) ngi, hu ht cc ri lon di truyn l ln (xem bng 1.5). i a s nhng ngi mc cc bnh ny thng c b m u bnh thng v kiu hnh, nhng li mang gene bnh trng thi d hp.
Bng 1.5 Mt s ri lon n gene thuc nhim sc th thng ngi (phng theo Campbell v Reece 2001; Lewis 2003) Ri lon Cc ri lon ln Bch tng Ha x nang Galactosemia Gaucher Hemochromatosis Phenylketonuria Bnh hng cu lim (ng hp t) Bnh Tay-Sachs Cc ri lon tri Achondroplasia Bnh Alzheimer Bnh Huntington Hypercholesterolemia Khng dung np lactose Hi chng Marfan Cc triu chng (v nguy c mc phi) Thiu sc t da, tc v mt (1/22.000) Tha cht nhy phi, dch tiu ha, gan Tch ly galactose cc m; tr n; tn thng mt v gan. (1/100.000; x l bng king galactose). Gan v lch sng phng, thiu mu, xut huyt ni C th gi li st; nguy c ly nhim cao, tn thng gan, tim v ty , tha sc t da Tch ly phenylalanyl trong mu; thiu sc t da bnh thng; tr n, hn m, cht tui nh (1/10.000) Tn thng lch v nhiu c quan; nguy c ly nhim cao (1/500 ngi M gc chu Phi; ng tri) Tch ly lipid trong cc t bo no gy suy thoi thn kinh; m mu; cht thi th u Ln vi t chi ngn, u v thn bnh thng Thoi ha tm thn; thng xy ra mun mng Thoi ha tm thn v cc c ng mt kim sot Tha cholesterol trong mu; bnh tim; 1/500 l d hp Khng c kh nng phn gii ng lactose, gy ra tnh trng chut rt sau khi n loi ng ny T chi di nh vn, ngc lm, hng thy tinh th, cc ngn tay mnh khnh, ng mch ch suy yu

Bnh thn a nang Tt nhiu ngn
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Cc bng trong cc qu thn, ur trong mu, huyt p cao, au bng di Tha cc ngn tay, ngn chn
Hai bnh ln in hnh l bch tng v ha x nang. Nhng ngi b bch tng (albino) l do thiu ht sc t melanin, nn da d trng bch, tc v trng en ca mt tr nn nht khc thng (hnh 1.5)... Ha x nang (cystic fibrosis) l mt bnh di truyn gy cht ph bin nht M (USA). Bnh ln ny ph bin nht nhng ngi M da trng gc Capca (Caucasians), vi tn s chung l 1/1.800, ngha l trung bnh c 25 ngi c mt ngi mang allele ln ny (Campbell v Reece 2001) hay i vi tr s sinh l 1/2.500 (Weaver v Hedrick 1997). Ngi b bnh ny c c im l tit ra mt lng d tha cht nhy dy phi, ty v cc c quan khc. Cc cht nhy ny c th lm nhiu lon s th, tiu ha v chc nng ca gan v lm cho ngi bnh ri vo nguy c vim phi v cc bnh truyn nhim khc. Nu khng c iu tr, hu ht tr em mc bnh ny s b cht tui ln nm. Theo thng k ca McKusick nm 1994, c 1.730 mc cho cc locus ln. 2. Cc tnh trng tri (dominant traits) Mc d hu ht cc allele c hi l allele ln, nhng mt s cc ri lon ngi l do cc allele tri (xem bng 1.5). Trong s c mt vi allele khng gy cht, chng hn nh tt tha cc ngn tay v chn (hnh 1.5), hoc c mng da gia cc ngn tay v chn. Cc tnh trng nh c tn nhang, di tai thng cng nh cc kh nng un li hnh ng v gp ngc li ln trn u do cc gene tri n khc nhau kim sot. Theo thng k ca McKusick nm 1994, c 4.458 mc cho cc locus tri.
Hnh 1.5 Mt s v d v di truyn Mendel ngi.

T tri sang l bch tng, tt nhiu ngn v dng ln ph bin (achondroplasia).
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Th d in hnh v ri lon tri nghim trng l dng ln ph bin do thoi ha sn gi l achondroplasia (hnh 1.5), u v thn mnh pht trin bnh thng nhng tay chn ngn mt cch bt thng; t l mc bnh ny l khong 1 trn 25.000 ngi. Ch nhng ngi d hp t mi b ri lon ny; cn kiu gen ng hp t tri gy cht phi. Trng hp khc l bnh Huntington (Huntington's disease), mt dng ri lon do s suy thoi ca h thn kinh thng xy ra t sau tui trung nin. Khi bnh tin trin, n lm cho cc c ng trn mi phn ca c th mt kh nng kim sot. S mt mt cc t bo no dn ti mt tr nh v kh nng suy xt, gp phn y nhanh s suy thoi. Cui cng, mt lun cc k nng vn ng lm cho khng nut v ni nng c. Ci cht thng xy ra sau khi cc triu chng bt u biu hin khong 10-20 nm (Campbell v Reece 2001).
VI. L thuyt xc sut trong d on v phn tch di truyn hc

hiu r cc pht hin ca Mendel v cc nguyn l ca di truyn hc ni chung, cng nh vn dng cc kin thc ny mt cch c hiu qu vo trong hc tp v thc tin i sng-sn xut, chng ta cn nm vng mt vi khi nim v nguyn l xc sut c bn sau y. 1. Mt s khi nim v tnh cht c bn ca xc sut (1) Mt cch n gin, xc sut (probability) c nh ngha bng s ln xy ra mt bin c hay s kin (event) c th chia cho tng s c may m bin c c th xy ra. Nu ta k hiu xc sut ca bin c A l P(A), m l s ln xut hin ca A v n l tng s php th hay ton b s kh nng c th c, khi : P(A) = m / n; trong 0 m n, v n > 0. Nh vy, 0 P(A) 1. (2) Php th l vic thc hin mt nhm cc iu kin xc nh, v d mt th nghim tung ng xu hay gieo ht xc xc hoc mt php lai c th. Cc kt cc khc nhau c th c t php th gi l cc bin c, c k hiu bng cc ch ci in hoa A, B, C...V d: Khi tung mt ng xu, s kin xy ra ch c th l mt sp (S) hoc nga (N) vi xc sut tng ng l 0,5. Tng t, kiu gene d hp Aa c th to ra hai loi giao t mang A v a vi xc sut ngang nhau l 0,5 trong khi cc kiu gene ng hp nh aa chng hn ch cho mt loi giao t mang a; v vy i con ca php lai phn tch Aa aa c t l k vng l 0,5 Aa : 0,5 aa. Khi thc hin php th c th xut hin mt trong cc loi bin c sau: -Bin c ngu nhin (A) l s kin c th xy ra nhng cng c th khng, 0 P(A) 1. -Bin c chc chn ( ) l s kin nht thit xy ra, P() = 1.
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-Bin c khng th c () l s kin nht thit khng xy ra, P() = 0. -Bin c xung khc: Hai bin c A v B gi l i xung khc vi nhau nu tch ca chng l mt bin c khng th c: AB = P(AB) = 0 v P(AB) = P(A) + P(B). -Bin c i lp: "khng A" () c gi l bin c i lp ca bin c A khi = \ A v A= . Khi P() = 1 P(A). -Nhm y cc bin c hay khng gian bin c s cp () l tp hp ton b cc bin c s cp () ca mt php th m khi c thc hin th nht thit mt trong chng phi xy ra, v c hin tng xung khc tng i. V d, dy cc bin c B1, B2,..., Bn lp thnh mt nhm y nu tho mn hai iu kin: (i) Tng ca chng l mt bin c chc chn: B1 B2 ... Bn = ; v (ii) Chng xung khc tng i mt: BiBj = ; ij (i, j = 1,2,...,n). 2. Mt s nguyn l xc sut c bn 2.1. Quy tc cng Quy tc cng pht biu rng: xc sut kt hp ca hai (hoc nhiu) s kin xung khc tng i xy ra l tng cc xc sut ring r ca chng. (i) Vi A, B l hai bin c bt k, ta c P(AB) = P(A) + P(B) P(AB) (ii) Nu A v B l hai bin c xung khc, th: P(AB) = P(A) + P(B) (iii) H qu: Nu l bin c i lp ca A, th P() = 1 P(A). V d, vi kiu gene Rr, nu ta k hiu xc sut ca loi giao t mang allele R l P(R) v ca loi giao t mang allele a l P(r), theo l thuyt ta c: P(R hoc r) = P(R) + P(r) = 1/2 + 1/2 = 1. 2.2. Xc sut iu kin nh ngha: Nu A, B l hai bin c bt k v P(A) > 0, th xc sut iu kin ca bin c B vi iu kin bin c A xy ra l: P(B/A) = P(AB) : P(A). V d: T kt qu th nghim lai gia hai th u thun chng ht vng v ht xanh ca Mendel, hy tm xc sut mt cy u ht vng F2 l th d hp (Vv). Bng lp lun thng thng ta thy trong s bn kiu t hp F2 c 3 kiu t hp cho kiu hnh ht vng (V-) nhng ch c 2 kiu l d hp (Vv). V vy xc sut cn tm l 2/3. Nu gii theo nh ngha xc sut, ta k hiu: A l s kin ht vng F2 v B l s kin ht vng d hp. Theo l thuyt, F2 c 4 s kin ng kh nng vi t l l 1VV: 2Vv: 1vv. y P(A) = 3/4 v P(A.B) = P(B) = 2/4 P(B/A) = P(AB) : P(A) = 2/4 : 3/4 = 2/3.
B B
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2.3. Quy tc nhn T nh ngha v xc sut iu kin, ta i n nh ngha v hai bin c c lp nh sau: Hai bin c A v B c gi l c lp vi nhau nu P(B/A) = P(B) hoc P(A/B) = P(A). Ngha l s xy ra hay khng xy ra ca bin c ny khng nh hng n s xy ra ca bin c kia. Khi , quy tc nhn c pht biu nh sau: Xc sut trng hp ca c hai bin c c lp bng tch cc xc sut ring r ca chng. Ngha l, nu A v B l cc bin c c lp th: P(AB) = P(A).P(B) V d: Khi gieo mt ng xu hai ln lin tip hoc gieo hai ng xu cn i v ng cht cng mt lt th c 4 kh nng xy ra l (SS), (SN), (NS), (NN) vi xc sut ngang nhau. Tht vy, xc sut xut hin c hai mt sp hoc nga l: P(S,S) = P(N,N) = 1/2 1/2 = 1/4; cn xc sut xut hin mt mt sp v mt mt nga l P(S,N) = 2 1/2 1/2 = 1/2; v tng cc xc sut ca tt c cc s kin ng nhin bng 1. T y ta c th xy dng c s ca "php th c lp", nguyn tc gii bi ton lai hai tnh v cc phng php biu din kt qu lai nh sau: (i) Mnh "php th c lp" di y c thit lp da trn quy tc nhn xc sut, ch yu dng kim tra xem quy lut di truyn chi phi ng thi c hai tnh trng no trong mt php lai l c lp hay lin kt (nu l lin kt th lin kt hon ton hay khng hon ton), hoc mt tnh trng no c lin kt vi gii tnh hay khng. Mnh ny c pht biu nh sau: Nu cc gene phn ly c lp v t hp t do, th t l phn ly ng thi ca c hai tnh trng bng tch cc t l phn ly ring r ca cc tnh trng , v ngc li. V d 1: Kt qu ca mt php lai hai tnh c t l phn ly l 3:3:1:1 v cc t l ca tng tnh l 3:1 v 1:1. Ta ni rng cc tnh trng tun theo quy lut phn ly c lp, v: 3:3:1:1 = (3:1)(1:1). V d 2: Kt qu s t th tinh ca mt c th F1(hoc tp giao gia cc c th F1 sinh ra t cc b m thun chng khc nhau v tng cp gene allele) v mt tnh trng no m c t l phn ly l 9:3:3:1 hoc l mt bin dng ca cng thc phn ly . Ta c th khng nh rng tnh trng ny tun theo quy lut tng tc gia cc gene khng allele phn ly c lp. Tht vy, t kt qu trn cho thy i lai c 9+3+3+1 = 16 kiu t hp vi t l ngang nhau, chng t (cc) c th F1 cho t nht 4 loi giao t vi t l tng ng, ngha l d hp t nht 2 cp gene phn ly c lp. Ngha l tnh trng tun theo quy lut tng tc gene. (ii) H qu: Nu tch cc t l phn ly ring r ca cc tnh trng khc vi t l phn ly ng thi ca c hai tnh, chng t cc tnh trng tun
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theo quy lut di truyn lin kt. V d: Kt qu ca mt php lai cho thy t l phn ly ca c hai tnh l 1A-bb:2A-B-:1aabb hoc 3A-B-:1aabb, trong khi t l phn ly ca mi tnh trng vn l 3:1.Ta d dng thy rng tch (3:1)(3:1) 1:2:1 hoc 3:1, chng t cc tnh trng ny tun theo quy lut lin kt hon ton, v kiu gene ca b m chng i vi trng hp u l Ab/aB Ab/aB hoc AB/ab Ab/aB, v trng hp sau l AB/ab AB/ab. (iii) Nguyn tc tng qut gii mt bi ton lai hai hoc nhiu tnh T nhng iu trnh by trn y cho thy rng thc ra ton lai gm hai dng tng qut: lai mt tnh v lai nhiu tnh, v vic gii mt bi ton lai hai tnh tr ln ni chung c thc hin theo ba bc chnh: Bc 1: Tch xt quy lut di truyn ca tng tnh trng; Bc 2: Gp xt quy lut di truyn ca hai tnh hoc tng hai tnh trng mt ( xc nh xem chng tun theo quy lut no, c lp hay lin kt); v Bc 3: Thit lp (cc) s lai kim chng (xem "Php gii ton lai", Hoa hc tr s 362 tr.40-41). Nh vy, ci kh ca cc bi ton lai thc ra l dng lai mt tnh, v ci kh nht ca dng ton ny l xc nh cho c quy lut chi phi tnh trng v cc kiu quan h c th c gia cc gene allele cng nh gia cc gene khng allele vi nhau. T mi c th xc nh cc kiu gene, kiu hnh, v cui cng l kim chng cc kt qu bng s lai thch hp. (iv) Cc phng php biu din kt qu lai: Bi v bt c mt mnh quy lut no cng l "mnh ko theo" v c tnh xc sut (ngha l cha ng kh nng tin on tim tng), cho nn vic tnh ton v biu din cc kt qu k vng ca bt k mt php lai no r rng l u da trn c s xc sut m ch yu l quy tc nhn. C nhiu cch khc nhau trong biu din kt qu ca cc php lai. Chng hn, trnh by di dng khung Punnett, vn l phng php kinh in ca di truyn hc. Mt phng php thng dng khc l phng php phn nhnh (forked-line approach), Phng php ny c th dng tm xc sut ca cc kiu i con i vi nhiu gene phn ly c lp; n cng rt tin dng khi xc nh thnh phn allele v xc sut ca cc loi giao t t mt kiu gene c th. Ngoi ra, tu trng hp c th s dng trc tip quy tc nhn (product rule). 2.4. Cng thc xc sut nh thc n gin, ta xt php th ng xu gm hai s kin i lp l mt sp (S) v mt nga (N), vi cc xc sut tng ng l p v q, trong q = 1-p. Gi s trong n php th c lp c tin hnh, s kin S xut hin k ln v s kin N l n-k. tnh cc xc sut ny ta phi s dng cng thc xc sut nh thc (binomial probability) sau y:
37
Pn(k) = Ckn .pk.qn-k ; k = 0, 1, 2,..., n. trong , Ckn l h s nh thc; y Ckn = n!/ k!(n-k)! Lu : Dy n php th ny cn gi l dy php th Bernoulli, v n tho mn ba iu kin: (1) N l dy c lp; (2) Trong mi php th, khng gian bin c s cp ch c hai bin c s cp i lp l S v N; v (3) Xc sut ca mt bin c S hoc N l khng thay i trong mi php th, P(S) = p v P(N) = q = 1 p. V d: Gieo mt ng xu lin tip ba ln, c th xy ra 23 = 8 kh nng c lp v th t trong 4 nhm sau y:
3 mt sp SSS 2 sp v 1 nga SSN SNS NSS 1 sp v 2 nga SNN NSN NNS 3 mt nga NNN
Vi gi thit p = q =1/2, da vo cng thc xc sut nh thc ta d dng tnh c xc sut ca mi trng hp trn nh sau: P(3 mt sp) = 1(1/2)3(1/2)0 = 1/8 P(3 mt nga) = 1(1/2)0(1/2)3 = 1/8 P(2 sp 1 nga) = 3(1/2)2(1/2)1 = 3/8 P(1 sp 2 nga) = 3(1/2)1(1/2)2 = 3/8 2.5. Cng thc xc sut ton phn Gi s dy B1,B2,...,Bn l mt nhm y cc bin c, ngha l chng c hp l mt s kin tt yu (B1B2...Bn = ) v gm tng i xung khc (BiBj = , vi i j; i,j = 1, 2,...,n); v gi A l mt bin c bt k. Khi : A = A = (B1B2...Bn ) A = (B1 A) (B2 A) ... (Bn A) p dng cc nh l cng v nhn, ta c cng thc xc sut ton phn P(A) = P(B1).P(A/B1) + P(B2).P(A/B2) +...+ P(Bn).P(A/Bn) = P(Bi).P(A/Bi) ; vi i = 1,..,n. 2.6. Cng thc Bayes Vi gi thit nh trn cng thm mt iu kin mi l php th c tin hnh v kt qu l s kin A xy ra. V cc Bi (i= 1, 2,...,n) lp thnh mt nhm y nn cng vi A phi c ch mt s kin Bk no xy ra. Vy khi A xut hin th s kin no trong s cc Bi c nhiu kh nng xy ra hn c? Gii p cu hi ny c ngha l cn tnh xc sut
B B B B B
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P(Bk/A). Vn dng nh l nhn xc sut, ta c: P(ABk) = P(A).P(Bk/A) = P(Bk).P(A/Bk) Suy ra: P(Bk/A) = P(Bk).P(A/Bk) : P(A) ; vi P(A) > 0. Thay P(A) t cng thc xc sut ton phn trn, ta c cng thc Bayes nh sau: P(Bk/A) = P(Bk).P(A/Bk) : P(Bi).P(A/ Bi) Trong cc P(Bi) c gi l cc xc sut tin nghim (v chng c xc nh trc khi tin hnh php th), cn P(Bk/A) c gi l xc sut hu nghim (v n ch c xc nh sau khi tin hnh php th). Cn lu rng, cc quy lut Mendel v thc cht l cc nh lut nhn xc sut, cho nn tin on cc nguy c di truyn cn s dng n cng thc Bayes, tc l nh l xc sut ca nguyn nhn. V bi ton t ra ch c th gii quyt trn vn mt khi bit c cc xc sut tin nghim.
B B
V d: Ly ngu nhin mt cy u ht vng F2 (trong th nghim ca Mendel) cho lai vi cy ht xanh, v th h lai nhn c tt c l 6 cy ht vng. Hy tm xc sut ca cy ht vng F2 em lai l th ng hp. (Quy c: Y- ht vng, y - ht xanh). Gii: Ta bit rng F2 c t l kiu gene l 1/4 YY : 1/2 Yy : 1/4 yy v t l kiu hnh l 3/4 vng : 1/4 xanh. V cy ht vng (Y-) c chn ngu nhin trong s cc cy ht vng F2 nn n c th l ng hp (YY) hoc d hp (Yy), vi xc sut tng ng l 1/3 hoc 2/3. Gi B1 - s kin "cy ht vng F2 ly ra l th ng hp" B2 - s kin "cy ht vng F2 ly ra l th d hp"; (B1B2 = ) v A l s kin "6 cy ht vng nhn c th h lai". Ta c cc xc sut tin nghim: P(B1) = 1/3 v P(B2) = 2/3 V cc xc sut iu kin: P(A/ B1) = 1 v P(A/ B2) = (1/2)6 = 1/64 Vy xc sut (hu nghim) cn tm l: P(B1/A) = P(B1).P(A/ B1) : [P(B1).P(A/ B1) + P(B2).P(A/ B2)] = (1/31) : [(1/31) + (2/31/64)] = 32/33 = 0,97.
B B B
Lu : Kt qu ny cho thy rng cy ht vng F2 (c ly ngu nhin lai phn tch) c kiu gene ng hp l hu nh chc chn (n 97%), vi mc = 0,05. Ni cch khc, ch vi 6 cy ht vng thi cng khng nh s ng tnh ca i con. n y hn l chng ta thy r cng thc Bayes c tm quan trng nh th no trong vic tin on nguyn nhn v nguy c di truyn, cng nh cho php l gii mt s tnh hung phc tp (xem Hong Trng Phn 1995, 2000b).
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VII. Phng php Khi-bnh phng (Chi-square method) trong nh gi ph hp gia cc s liu quan st v k vng
Ni chung, cc s liu thng k thu c t cc th nghim vn sai khc t nhiu so vi cc con s mang tnh cht l thuyt, tu thuc ch yu vo mu th nghim v phng php ly mu. Trong trng hp , chng ta bn khon khng r liu s gi nh "xp x" ca chng ta c tht chc chn khng? Hay ni theo ngn ng thng k, "gi thuyt tng ng H0 c chp nhn hay b bc b", ngha l kt qu thu c c tht nghim ng vi t l ca mt quy lut no hay khng? c c cu tr li rt ro cho vn ny ch c cch l s dng trc nghim Khi-bnh phng (2-test). y l mt cng c ton thng k thng dng cho php kim tra ph hp gia cc tr s thc t quan st c (observed, k hiu: O) v cc tr s l thuyt c k vng (expected, k hiu: E) ca mt gi thuyt hay phn phi thc nghim khoa hc no , hoc kim tra tnh c lp ca hai i lng ngu nhin. Nh ta c th rt ra quy lut, hoc hiu qu ca hai phng php th nghim no . ng v phng din thc hnh, phng php ny c tin hnh n gin nh sau: Bc 1: t gi thuyt tng ng H0 v sau tnh tr s 2 thc t da theo cng thc: 2 = [(Oi Ei)2/ Ei ] ; i= 1, 2,...,n. Bc 2: Tm tr s 2 l thuyt bng cch tra Bng cc gi tr ca phn phi 2 vi k bc t do. Thng thng ngi ta s dng mc xc sut sai lm P hay mc ngha = 0,05 v k = n 1; trong n l s lp kiu hnh v n cn tu trng hp c th. Mc = 0,05 thng c dng lm ranh gii phn chia gia min n nh chp nhn gi thuyt H0 v min n nh bc b gi thuyt H0. tin li, di y nu ra mt vi tr s 2 = 0,05 l thuyt thng dng ng vi mt s bc t do k nh sau: S bc t do (k) Gi tr
2 = 0,05
1 3,84
2 5,99
3 7,82
4 9,49
Bc 3: So snh cc gi tr 2 thc t v l thuyt. - Nu nh tr s 2 thc t nh hn tr s 2 l thuyt, tc l c mc xc sut P hay > 0,05, gi thuyt H0 c chp nhn. Ngha l kt qu thu c ph hp vi t l c gi nh. - Ngc li, nu nh tr s 2 thc t ln hn hoc bng tr s 2 l thuyt, tc l c mc xc sut P hay 0,05, gi thuyt H0 b bc b. Ngha l kt qu thu c khng ph hp vi t l c gi nh.
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V d: Trong th nghim lai mt tnh ca Mendel, F2 thu c 705 hoa tm v 224 hoa trng. Trn nguyn tc, vi hai kiu hnh F2 khin ta c th ngh ti mt s t l l thuyt gn vi n nh 2:1, 3:1 hoc 9:7. Nhng y t l "tm : trng" thc t l 3,15:1 nn t l k vng hp l hn c l 3:1. chnh l gi thuyt H0 cn kim tra. By gi ta c th tnh ton gi tr 2 thc t nh sau:
Kiu hnh Hoa tm Hoa trng Tng S quan st (Oi) 705 224 929 S k vng (Ei) 3/4 929 = 696,75 1/4 929 = 232,25 929 (Oi Ei)2/ Ei 0,098 0,293 2 = 0,391
Bng cch tra bng cc gi tr ca phn phi 2 = 0,05 vi k = 21= 1 bc t do, ta tm c tr s 2 l thuyt l 3,84. V tr s 2 thc t (0,391) nh hn tr s 2 l thuyt (3,84) rt nhiu, nn gi thuyt H0 hon ton c chp nhn. Ngha l kt qu th nghim trn ph hp mt cch st sao vi t l 3:1. iu c ngha rng s sai khc gia cc s liu thc v cc con s l thuyt tng ng l rt khng ng k, khng c ngha v phng din thng k.
Cu hi v Bi tp
1. i tng v phng php th nghim ca Mendel c nhng c im c o no m t Mendel khm ph ra cc quy lut di truyn c s u tin, t nn mng cho s ra i ca di truyn hc? 2. Khi lai gia chut en v chut nu nhn c F1 ton chut en. Sau cho cc chut ny tp giao vi nhau thu c F2 gm 53 en v 17 nu. Hy gii thch kt qu, vit cc s lai v tnh xc sut ca mi trng hp phn ly kiu hnh c th c F2 (bit rng s chut F2 thu c t mi t hp lai ca cc chut F1 trung bnh l bn con). 3. T kt qu lai cu sau y, hy xc nh kiu gene ca mi c th. Php lai i con Trng-1 x trng-2 6 trng : 1 en Trng-1 x trng-3 5 trng Trng-1 x en-1 3 trng : 3 en 4. Cho mt rui gim mt nu cnh di giao phi vi mt rui gim mt cnh di. i con thu c gm 51 di, 53 nu di, 17 ngn v 18 nu ngn. Hy xc nh kiu gene ca cc rui gim b m.
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5. Da vo cc php lai v kt qa di y, hy bin lun ch ra cc quy lut di truyn v kiu gene ca cc b m trong mi php lai.
i con Php lai Vng, trn vng, trn Vng, nhn vng, nhn Xanh, trn vng, nhn Vng, trn 45 0 31 Vng, nhn 15 42 29 Xanh, trn 16 0 32 Xanh, nhn 5 15 30
6. Gi s thc hin php lai gia con mo lng ngn, tha, c m trng (llddSs) v mo lng di, rm, c m trng (LlDdSs). Hy trnh by cc phng php nhn xc sut n gin v s phn nhnh tnh t l k vng ca mi mt mo con c kiu hnh sau: lng ngn, tha, khng c m trng; lng ngn, rm, c m trng; v lng di, rm, c m trng. 7. rui gim, mt (se+) l tri so vi mt mu nu ti (se; vit tt ca sepia). Cho hai rui gim mt d hp lai vi nhau, v ly mt con mt i con cho lai tr li vi mt b m mt . C may i con php lai ngc xut hin rui gim mt mu nu ti l bao nhiu? 8. khng nh cc nguyn l di truyn ca Mendel, cc nh nghin cu lp li cc th nghim u H Lan. Ring cc th nghim v mu sc ht, F2 Correns thu dc 1.394 vng v 453 xanh, trong khi Tschermak quan st thy c 3.580 vng v 1.190 xanh. Cc tr s ny c ph hp vi nguyn l phn ly khng? Bn hy chng minh iu bng cng c ton thng k thch hp. 9. Gi s ngi chng b bch tng, ngi v v cc con ca h (mt trai v hai gi) u bnh thng. Khi ln ln, chng lp gia nh vi nhng ngi cng khng mc bnh ny v mi gia nh ny u c mt trai v mt gi bnh thng. Gi s xy ra s kt hn ng huyt gia hai a chu c quan h cu c rut, khi xc sut sinh ra mt trai u lng mc bnh ny l bao nhiu? Hy v s ph h ni trn v gii thch. 10. Gi s bn c mt cy u H Lan thn cao cha r kiu gene. Bn em cy ny lai phn tch vi cy thn thp hoc cho n t th phn v thu c tt c by cy u c thn cao. Hi xc sut cy thn cao em lai kim tra c kiu gene ng hp l bao nhiu?
Ti liu Tham kho

Ting Vit Nguyn Ngc King 1996. Thng k hc trong Nghin cu Khoa hc.
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NXB Gio Dc, H Ni. Phm Vn Kiu 1998. L thuyt Xc sut v Thng k Ton hc. NXB Khoa hc v K thut, H Ni. Hong Trng Phn 1995. nh gi hin tng ng tnh trong lai phn tch nh cng thc Bayes. Thng tin Khoa hc v Gio dc s 11/1995, tr. 133-138, Trng HSP Hu. Hong Trng Phn 2000a. Php gii ton lai. Hoa hc tr s 362, tr.40-41. Hong Trng Phn 2000b. Cng thc Bayes v php tnh gn ng trong phn tch di truyn hc. Thng bo Khoa hc s 4/2000, tr. 65-76, Trng HSP H Ni. Ting Anh Brooker RJ. 1999. Genetics - Analysis and Principles. Benjamin/ Cummings Publishing Company, Inc., Menlo Park, CA. Campbell NA, Reece JB. 2001. Essential Biology. Benjamin/Cummings, an imprint of Addison Wesley Longman, Inc, San Francisco, CA. Hartl DL, Freifelder D, Snyder LA. 1988. Basic Genetics. Jones and Bartlett Publishers, Inc, Boston - Portola Valley. Lewis R. 2003. Human Genetics: Concepts and Applications. 5th ed, McGraw-Hill, Inc, NY. Online Mendelian Inheritance in Man (OMIMTM): <http://www.ncbi.nlm.nih.gov/Omim/mimstats.html> Russell PJ. 2000. Fundamentals of Genetics. 2nd ed, Benjamin/ Cummings Publishing Company, Inc, Menlo Park, CA. Suzuki DT, Griffiths AJF, Miller JH, Lewontin RC. 1989. An Introduction to Genetic Analysis. 4th ed, W-H Freeman and Company, New York. Tamarin RH. 1999. Principles of Genetics. 6th ed, McGraw-Hill, Inc, NY. Weaver RF and Hedrick PW. 1997. Genetics. 3rd ed, McGraw-Hill Companies, Inc. Wm.C.Brown Publishers, Dubuque, IA. Mt s trang web http://www.mhhe..com/lewisgenetics5 htttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=OMIM http://fpt.netspace.org/MendelWeb/Mendel.htm http://www.nord-rdb.com/~orphan

Chng 2
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M rng v p dng ca Di truyn hc Mendel

Nt c sc ca cc th nghim Mendel l s nht qun hon ton gia cc quan st v gi thuyt ca ng. Tuy nhin, k t sau khi cc cng trnh ca Mendel c khm ph li, ngi ta cn pht hin ra nhiu ngoi l suy rng t m hnh Mendel. Trong chng ny, chng ta s tm hiu cc vn sau: (i) S khc nhau gia cc kiu tri hon ton, tri khng hon ton v ng tri; (ii) Tc ng ca gene gy cht; (iii) Hin tng a allele; (iv) Tnh a hiu ca gene; (v) Cc kiu di truyn do nhiu gene cng tc ng ln mt tnh trng - tng tc gene; (vi) C s di truyn ca cc tnh trng s lng. Cc kiu di truyn lin kt v lin kt vi gii tnh tuy cng c xem nh l s m rng ca cc nguyn l Mendel, nhng s c trnh by trong mt chng ring, chng 4.
I. Cc kiu quan h gia cc gene allele i vi mt tnh trng

1. Cc kiu tri hon ton, khng hon ton v ng tri K t sau nm 1900, ngi ta cn pht hin thm mt s trng hp tri khc nhau, b sung cho t l 3 tri :1 ln ca Mendel. 1.1. Tri hon ton (complete dominance) y l trng hp di truyn tri-ln Mendel. Trong hu ht cc trng hp, allele bnh thng (hay kiu di) tri hon ton so vi cc allele t bin. iu ny c th l gii da trn c s di truyn sinh ha ch, allele tri cho sn phm protein hot ng chc nng bnh thng trong khi allele t bin khng to ra c sn phm c hot tnh. Do cc c th ng hp v allele ln khng hon thnh c con ng chuyn ha c lin quan n gene ny. ngi, l trng hp ca cc allele t bin ln gy bch tng, bnh phenylxtn-niu (phenylketonuria = PKU)... Tuy nhin, mt s trng hp, allele t bin tri hn kiu di; ngha l allele kiu di l ln. V d: ngi, kiu ln ph bin do khng to c sn l tri, cho nn cc th d hp biu hin kiu hnh t bin. 1.2. Tri khng hon ton (incomplete dominance) Khi lai gia hai ging hoa bn gi (four-o'clock; Mirabilis jalapa) thun chng c hoa mu v hoa mu trng, Carl Correns thu c tt c cc cy F1 c hoa mu hng, kiu hnh trung gian gia hai b m. Sau khi cho cc cy F1 t th phn, t l kiu hnh F2 l 1 : 2 hng : 1 trng. Mc d t l kiu hnh ny c hi lch so vi ca Mendel, nhng
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thc t n tng ng vi t l kiu gene 1:2:1 (hnh 2.1). Nu s dng quy c gene A- l tri khng han ton so vi a- trng, ta c s lai sau: Ptc Hoa (AA) Hoa trng (aa) Aa (Hoa hng) F1 F2 T l kiu gene AA : Aa : aa T l kiu hnh : hng : trng
tri ng hp t
d hp t
ln ng hp t
Hnh 2.1 S di truyn trung gian i vi mu sc hoa nhiu thc vt.
Bi kiu hnh ca th d hp l trung gian gia hai th ng hp, v vy ta c th l gii trn phng din sinh ha rng hm lng sn phm tch ly do mt allele tri kim sot l khng th hin kiu hnh mu nh trong trng hp c mt c hai allelele tri. 1.3. ng tri (codominance) ng tri l hin tng c hai allele khc nhau trong mt th d hp cng biu hin ra cc sn phm c hot tnh khc nhau trong t bo. Cc allele nh th c gi l cc allele ng tri. in hnh l trng hp nhm mu AB ca h nhm mu ABO (hnh 2.2; xem gii thch mc 3 bn di) v nhm mu MN ca h nhm mu M-N ngi.
Hnh 2.2 Kiu hnh cc nhm mu A, AB v B. ( y cho thy s ng tri nhm mu AB. Nhm mu O khng c khng nguyn no).
H nhm mu M-N (do mt locus thuc nhim sc th thng kim sot) c hai allele LM v LN. Nh th, trong mt qun th s c ba kiu gene LMLM, LMLN v LNLN (c th vit gn l MM, MN v NN) tng ng vi ba kiu hnh hay nhm mu l M, MN v N. Nu cho rng cc php hn phi thun nghch l tng ng, th c th c su kiu hn phi vi cc t l kiu gene k vng i con c cho bng 2.1.

Bng 2.1 Cc t l k vng i con i vi h nhm mu M-N B m i con
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L L
M M
LMLN
LNLN
LMLM LMLM 1 M M M N L L L L M M N N L L L L 1 M N M N L L L L M N N N L L L L N N N N L L L L 1 Mt v d khc l allele ln gy bnh hng cu hnh lim. nhng ngi d hp t v allele ny (HbAHbS), c hai allele u c biu hin v cc t bo mu ca h cha c hemoglobin bnh thng v bt thng. 2. Tc ng ca gene gy cht (lethal) Cc allele gy cht l nhng t bin c th tri hoc ln lm gim sc sng hoc gy cht i vi cc c th mang n v do , lm bin i t l 3:1 ca Mendel. Nhiu gene c cc allele nh hng ln t l cht ch khng gy cht; cc allele ny c gi l cc allele c hi (deleterious).
Hnh 2.3 Bin i mu lng chut.
Hnh 2.4 Mo Manx khng ui.
Ni chung, cc allele gy cht thng l ln v gy cht cc th ng hp. V d, t bin bch tng thc vt lm cho cy cht giai on non v khng c dip lc quang hp. Bnh thiu mu hng cu hnh lim ngi (xem mc II) c th gy cht vi t l ng k tui trng thnh khi allele t bin ln ny trng thi ng hp. Tuy nhin, mt s allele gy cht l nhng t bin tri. in hnh l th nghim lai v mu sc lng chut ca Lucien Cunot nm 1904. Khi lai gia hai chut thn vng (allele vng l tri; Hnh 2.3), ng thu c t l xp x 2 vng : 1 kiu di. Mt khc, khi lai gia cc chut vng vi chut kiu di (mu agouti), ng thy rng i con c t l xp x 1:1.
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Cunot kt lun rng tt c cc chut vng u l nhng th d hp, cn cc th ng hp v allele vng u b cht giai on phi. B m Yy (vng) Yy (vng) YY : Yy : yy i con
(cht) 2 vng : 1 agouti
Hin tng "khng ui" mo Manx (c pht hin u tin o Manx nm 1935; Hnh 2.4) l mt tnh trng khc gy ra bi mt allele t bin tri M; n c hiu qu tri cc th d hp v gy cht cc th ng hp. V vy khi lai gia cc mo Manx (Mm Mm) bao gi cng thu c mo Manx khng ui v c ui bnh thng. 3. Hin tng a allele (multiple allelelism) Trn thc t, mi mt gene khng ch c hai allele m c th c nhiu hn hai allele, gi l a allele. Cc allele l nhng trng thi cu trc khc nhau ca cng mt gene pht sinh do t bin (xem chng 8). Ni chung, nu mt gene trn nhim sc th thng (autosome) c n allele khc nhau, th trong qun th c th c n(n+1)/2 kiu gene, trong c n kiu ng hp v n(n 1)/2 kiu d hp (xem bng 2.2). y chnh l c s ca hin tng a hnh (polymorphism) quan st c trong cc qun th t nhin.
Bng 2.2 Mi quan h s lng gia cc allele v cc kiu gene S allele 1 2 3 4 ... n S th ng hp 1 2 3 4 ... n S th d hp 0 1 3 6 ... Tng s kiu gene 1 3 6 10 ...
n(n 1)/2
n(n+1)/2
V d: S di truyn h nhm mu ABO ngi c kim sot bi mt gen autosome c ba allele chnh l IA, IB v IO; trong IO l ln, cn cc allele IA v IB l ng tri. Trong mt qun th, ni chung c su kiu gen tng ng vi bn kiu hnh hay nhm mu sau y: Kiu hnh A B AB O A A A O B B B O A B O O Kiu gene I I ,I I I I , I I I I I By gi ta hy tm hiu c s di truyn min dch ca h nhm mu ABO ny. S khc nhau gia cc nhm mu l do s c mt ca cc loi khng nguyn v khng th . Cc khng nguyn (antigen) y l nhng phn t kt hp protein-ng bm trn b mt ca cc t bo hng cu v
47
chng xc nh tn nhm mu tng ng. Cc c th mang allele IA v/hoc allele IB c khng nguyn tng ng A v/hoc B; cn allele IO khng to c bt k khng nguyn no (nn nhng ngi ng hp v allele ln ny c kiu hnh O, ngha l "khng" c khng nguyn A hoc B no c trn b mt cc t bo hng cu). Cc khng th (antibody) l nhng protein do h thng min dch to ra vi mt s lng ln nhm p ng vi cc khng nguyn c th t bn ngoi. C th, trong huyt thanh nhng ngi mang nhm mu O thy c c hai loi khng th khng-A v khng-B gi l v ; nhng ngi nhm mu AB khng c bt k loi khng th no; cn nhng ngi nhm mu A v B ch c mt loi khng th tng ng l v . ngha lm sng ca nguyn tc truyn mu l ch khi truyn sai nhm mu s xy ra phn ng ngng kt cht ngi do phn ng gia khng nguyn b mt ca hng cu ngi cho (donor), m lng huyt thanh l khng ng k, vi khng th c trong huyt thanh ca ngi nhn (recipient). V th, nhng ngi c cng nhm mu th c th cho v nhn ca nhau. c bit, nhm mu O do thiu c hai loi khng nguyn A v B nn c th truyn cho mt ngi mang bt k nhm mu no; ngc li ngi mang nhm mu AB c th nhn bt k nhm mu no. Hin tng a allele tn ti ph bin trong cc qun th sinh vt khc nhau. V d, gene kim sot mu mt -trng rui gim gm mt chui 12 allele, vi tnh tri gim dn t kiu di cho n trng t bin ln (w) theo th t t tri sang phi v trn xung nh sau:
Allele Mu Allele Mu W+ di We eossin Wsat satsuma Wbl blood Wco coral Wap apricot Ww wine Wi ivory Wap3 apricot3 Wt tinged Wch cherry w white
Mt s gene ngi, chng hn nh cc gene i vi khng nguyn bch cu ngi HLA (human leukocyte antigen) xc nh cc khng nguyn trn b mt ca hu nh tt c cc t bo c th c nhiu allele. V d, gene HLA-B c nhiu hn 30 allele c xc nh khc nhau v mt khng nguyn trong mt s qun th. Kt qu ca s a dng ny l, trong mt qun th c rt nhiu kiu gene gene HLA-B (465 kiu gene khc nhau, vi 30 kiu ng hp v 435 kiu d hp), thc s to nn mt dy bin d kiu gene rng.
II. Tnh a hiu ca gene (pleiotropy)

Hin tng mt gene nh hng n hai hoc nhiu tnh trng c gi l tnh a hiu (pleiotropy). V d, trong cc th nghim u H Lan,
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Mendel lu rng gene kim sot mu hoa tm v trng cng nh hng ln mu sc ht (v xm hoc nu) v gy ra s c mt hoc khng c mt ca cc vt mu tm b l. Trong v d allele kim sot lng vng chut ni trn, ta thy rng n cn nh hng ln sc sng cc th d hp v gy cht cc th ng hp. Nhiu bnh di truyn ngi gy ra bi cc gen c tc dng a hiu. Chng hn, bnh phenylxtn-niu (phenylketonuria = PKU) xy ra cc c th ng hp v allele ln . Nhng ngi mc bnh ny thiu hn enzyme cn thit cho s chuyn ha bnh thng ca amino acid phenylalanine thnh sn phm sinh ha k tip. Khi so snh cc c th bnh thng v PKU vi nhau cho thy mc phenylalanine nhm bnh cao hn nhiu. Ngoi ra, nhng ngi bnh cn c mt s bin i khc nh: ch s IQ thp hn, u b hn, v tc hi nht hn. Tt c cc hiu qu a hiu ny c th hiu nh l hu qu ca s ri lon sinh ha c s. Chng hn, cc bnh nhn PKU c s tch ly mt c t trong u khin cho b no b tn thng v dn ti IQ thp hn, u b hn. Mt v d khc v tnh a hiu ngi l bnh hng cu hnh lim (sickle-cell disease). Vy tnh a hiu gii thch trng hp ny nh th no? Nhng ngi ng hp v allele t bin ln ny (HbSHbS) ch to ra cc phn t hemoglobin bt thng, khin cho tt c cc t bo hng cu c dng hnh lim, kch thc b, mu nht (Hnh 2.5). Cc t bo hnh lim nhanh chng b c th ph hy v gy ra s thiu mu v suy yu c th ni chung. Ngoi ra,
Hnh 2.5 Cc t bo hng cu bnh thng (trn) v dng hnh lim (di), vi phng i gn gp i ca hnh trn.
do hnh dng gc cnh m cc t bo hnh lim khng th vn chuyn trn tru trong mu v c xu hng tch t v gy tc nghn cc mao mch. Dng mu i n cc b phn c th b gim bt, gy st nh k, au n, v tn thng cc c quan khc nhau nh no b, lch, tim, thn...
III. Cc kiu tng tc gia cc gene khng allele

K t sau khi pht hin li cc nguyn l di truyn ca Mendel, xut hin nhiu cng trnh nghin cu trn cc i tng khc nhau, nh: b ng, u ngt, la m, ng, rui gim, g, chut, ch, nga...Mt s kt qu thu c t cc th nghim lai mt tnh cho thy t l phn ly khc vi cc t l 3:1 hoc 9:3:3:1. Cc trng hp cho t l phn ly 9:3:3:1
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hoc mt dng bin i ca n c l gii l do s tng tc qua li gia cc gene khng allele phn ly c lp. Tu trung c hai kiu tng tc gene, l: tng tc gia cc gen allele ( xt trn) v tng tc gia cc gen khng allele (thng c gi tt l tng tc gene). Do cc gene khng allele c th nm trn cng mt cp nhim sc th hoc trn cc cp nhim sc th tng ng khc nhau, cho nn v nguyn tc s c hai dng tng tc gene khng allele tng ng l tng tc c lp v tng tc lin kt; v dng sau phi l ch yu bi v s lng gene ca mt sinh vt nhiu hn s lng nhim sc th ca n rt nhiu. Tuy nhin, trn thc t, khi ni n s tng tc gia cc gene khng allele l ta mun ni ti s tng tc gia cc gene khng allele phn ly c lp. Lu : (1) S lng cc tnh trng hnh thnh nn mt c th sinh vt ni chung l nhiu hn s lng gene m ha protein c mt trong b gene ca n. Chng hn, theo c tnh gn y cho thy trong b gene ngi c khong 25.000 gene khc nhau trong khi c ti 50.000 loi protein thc hin tt c mi hot ng sng ca cc t bo. (2) Nh th, s biu hin ca a s tnh trng l kt qu ca s tng tc gia nhiu gene khc nhau trong qu trnh pht trin c th; v thc cht ca s tng tc l do s tng tc gia cc sn phm hot ng ca cc gen c bn cht protein hoc gia cc sn phm sinh ra t s xc tc ca cc enzyme. (3) Con ng chuyn ha mt cht c th trong t bo l mt chui gm nhiu phn ng sinh ha ni tip nhau m mi khu phn ng c xc tc bi mt enzyme (chng 7). Ngay c mt enzyme hay protein c cu trc bc bn cng gm mt s chui polypepetide thuc cc gene khc nhau to nn (chng 6). (4) cho n gin, thng thng ta gii thch cc kt qu tng tc bng hai gene nhng trn thc t, c th c nhiu hn hai gene cng quy nh mt tnh trng. Di y chng ta xt ba kiu tng tc gen chnh: Tng tc b tr, tng tc t ch v tng tc a phn cng gp. Cc v d nu ra di y l nhng th nghim kinh in ca di truyn hc. hiu c bn cht di truyn ca cc tnh trng , c bit l cc tnh trng mu sc, chng ta c gng lm sng t c ch sinh ha ca chng trong kh nng c th. 1. Tng tc b tr (complementary) Tng tc b tr l trng hp tng tc gene lm xut hin kiu hnh mi khi c mt ng thi cc gene khng allele trong mt kiu gene. Cc gene b tr c th l gene tri hoc gene ln ( trng thi ng hp). Tng tc kiu b tr biu hin di nhiu dng vi cc t l kiu hnh F2 khc nhau, nh 9:3:3:1; 9:6:1; 9:7.
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1.1. Tng tc b tr vi t l 9:3:3:1 V d kinh in cho trng hp ny l cc th nghim ca W.Bateson v R.C.Punnett v s di truyn hnh dng mo g. Khi lai gia cc ging g thun chng mo hnh hoa hng vi mo n (cn gi l mo hnh l) thu c F1 ton mo hoa hng, v sau khi cho tp giao F1 th F2 c t l phn ly 3 mo hoa hng : 1mo n. Tng t, khi lai gia cc ging g thun chng mo hnh ht u vi mo n, F1 gm tt c mo ht u v F2 phn ly 3 mo ht u : 1 mo n. Nhng khi lai gia hai ging g thun chng mo hoa hng v mo ht u, th F1 li thu c tt c c mo hnh qu c ch (hay ht h o) v t l phn ly F2 xp x 9 qu c ch : 3 hnh hoa hng : 3 hnh ht u : 1 mo n (hnh 2.6). Gii thch: Cc kt qu trong hai th nghim u cho thy cc dng mo hoa hng v ht u l tri so vi dng mo n. Kt qu sau cng cho thy F2 c 16 kiu t hp vi t l ngang nhau, trong khi F1 ng nht kiu gen (v b m thun chng); iu chng t F1 cho 4 loi giao t vi t l tng ng, ngha l d hp t v hai cp gene phn ly c lp. Suy ra tnh trng ny do hai gene khc nhau chi phi, ngha l tun theo quy lut tng tc gene. Mt khc, kiu hnh mi biu hin F1 v khong 9/16 F2 (ng vi s c mt ca c hai gene tri khng allele) phi l kt qu ca s tng tc gia cc gene tri khng allele theo kiu b tr. Quy c: Da vo t l phn ly kiu hnh F2 trong trng hp hai gene phn ly c lp v tn gi cc dng mo theo ting Anh (rose: hoa hng, pea: ht u), ta c
c ch: R-P-
hoa hng: R-pp
ht u: rrP-
mo n: rrpp
Hnh 2.6 Cc kiu mo c trng ca cc ging g khc nhau v cc kiu gene tng ng.
th quy c v kim chng n gin nh sau: R-P-: mo hnh qu c ch (do b tr gia cc gene tri R v P) R-pp: mo hnh hoa hng (do biu hin ca gene tri R) rrP-: mo hnh ht u (do biu hin ca gene tri P) rrpp: mo n (do khuyt c hai gene tri; kiu di) Kim chng: Ptc mo hoa hng (RRpp) mo ht u (rrPP) F1 mo qu c ch (RrPp)
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F1F1 = RrPp RrPp = (Rr Rr)(Pp Pp) F2 = (3R-:1rr)(3P-:1pp) = 9 R-P-: 3 R-pp : 3 rrP- : 1 rrpp = 9 c ch: 3 hoa hng : 3 ht u : 1 n Mt v d khc, l lai gia hai ging chut lng mu en vi mu v qu (cinnamon) c F1 ton chut lng mu xm (c dng "mui tiu" khi nhn gn, cn gi l agouti) v F2 cho t l 9 xm : 3 mu v qu : 3 en : 1 nu. Bn c th gii thch c s di truyn ca trng hp ny? 1.2. Tng tc b tr vi t l 9:7 V d: Th nghim ca Bateson v Punnett v s di truyn mu sc hoa cy u ngt (Lathyrus odoratus). T php lai gia hai ging hoa trng thun chng khc nhau, hai ng thu c F1 gm tt c cy lai c hoa mu ta (purple). Khi cho cc cy F1 t th phn F2 nhn c 382 hoa ta v 269 hoa trng; kt qu ny gn vi t l 9:7. Gii thch: Vi cch lp lun nh trn, ta d dng thy rng kiu hnh hoa ta l kt qu ca s tng tc b tr gia hai gene tri khng allele phn ly c lp. Trc khi i vo gii thch c s di truyn sinh ha ca cc kiu hnh, ta hy quy c v kim chng kt qu th nghim trn. Quy c: C-P- : hoa ta (do tc ng b tr gia cc gene tri C v P) C-pp, ccP-, ccpp : hoa trng (do khng c mt y c hai gene tri) Kim chng: Ptc ging hoa trng 1(CCpp) ging hoa trng 2 (ccPP) F1 hoa ta (CcPp) F1F1 = CcPp CcPp = (Cc Cc)(Pp Pp) F2 = (3C-:1cc)(3P-:1pp) = 9 C-P- : (3 C-pp + 3 ccP- + 1 ccpp) = 9 ta : 7 trng By gi ta th gii thch c s sinh ha ca cc kiu hnh. S hnh thnh mu hoa cy u ngt l kt qu ca s tng hp mt hp cht gi l anthocyanin; n xy ra thng qua mt chui cc khu chuyn ha c xc tc bi cc enzyme vn l sn phm ca cc gene. Nu nh bt k khu no trong qu trnh tng hp ny b gin on do vng mt ca mt enzyme hot ng th s hnh thnh mu sc khng xy ra. Mc d chi tit chnh xc ca s tng hp mu sc cy u ngt cn cha r, nhng c th minh ha bng m hnh tng qut hnh 2.7.

Kiu gene c cha C Kiu gene c cha P Enzyme (C) Enzyme (P) Cht tin thn Sn phm trung gian Anthocyanin
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Hnh 2.7 S minh ha mi quan h gia cc gene tri C v P trong qu trnh hnh thnh sc t anthocyanin cy u ngt.
i vi cc kiu gene c cha c hai gene tri C v P (C-P-), c y cc enzyme cn thit cho vic to ra anthocyanin v do vy hoa c mu ta. Trong khi , kiu gene cha cc (ccP- hay ccpp), enzyme th nht khng c to ra hay khng c hot tnh; hu qu l, phn ng to sn phm trung gian khng thc hin c. Tng t, kiu gene cha pp (Cpp hoc ccpp) th phn ng th hai bin i cht trung gian thnh anthocyanin b dng li, v thiu hn mt enzyme tng ng. Ni cch khc, nu nh kiu gen c mang cp allele hoc l cc hoc l pp th con ng tng hp b gin on v cc sc t khng c to ra, v kt qu l hoa mu trng. 1.2. Tng tc b tr vi t l 9:6:1 V d: S di truyn hnh dng qu b ng. Khi lai hai ging b ng thun chng qu trn khc ngun gc vi nhau, F1 xut hin ton dng qu dt, v F2 c s phn ly kiu hnh xp x 9 dt : 6 trn : 1 di. Gii thch: Kt qu ny c th c gii thch theo mt trong hai cch sau: b tr gia cc gene tri hoc b tr gia cc gene ln. n gin, di y ta gii thch theo cch u, da trn quy c sau y: Quy c: D-F- : qu dt (do tng tc b tr gia cc gene D v F) D-ff v ddF- : qu trn (ch c mt trong hai gene tri D, F) ddff : qu di (do khuyt ng thi c hai gene tri) Kim chng: qu trn-1 (DDff) qu trn-2 (ddFF) Ptc F1 qu dt (DdFf) F1F1 = DdFf DdFf = (Dd Dd)(Ff Ff) F2 = (3D-:1dd)(3F-:1ff) = 9 D-F-: (3 D-ff + 3 ddF-) : 1 ddff = 9 qu dt : 6 qu trn : 1 qu di 2. Tng tc t ch (epistasis) Tng tc t ch l hin tng mt gene ny km hm s biu hin ca mt gene khc khng allele vi n. Gene t ch c th l tri hoc ln.
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2.1. t ch do gene ln vi t l 9:3:4 V d: S di truyn mu sc lng chut (hnh 2.8). Khi lai gia hai dng chut thun chng lng nu v bch tng, F1 xut hin ton chut lng en; v khi cho cc chut F1 tp giao vi nhau th F2 c s phn ly gn vi t l 9 en : 3 nu : 4 bch tng. gii thch kt qu ny, ta da vo bin lun nh v d u tin (mc III-1.1) v quy c sau y: Quy c: B-C- : en; bbC- : nu; B-cc v bbcc : bch tng Gii thch: T quy c ny, ta thy rng allele c khi trng thi ng hp (cc) c ch s biu hin ca B- v bb khin cho cc kiu B-cc v bbcc khng c sc t (bch tng, albino). i vi hai kiu hnh cn li c th gii thch theo mt trong hai cch sau: (1) Allele C l t bin tri, nn mt kh nng t ch v bn thn n khng to mu; khi allele tri B quy nh mu en l tri so vi allele b quy nh mu nu khi n trng thi ng hp; kt qu l B-C- c kiu hnh lng en v bbC- cho kiu hnh lng nu. Nh th y khng xy ra s tng tc b tr gia cc gene tri B v C. Cch l gii ny r rng l hp l. (2) Allele C quy nh mu nu v B l gene to mu, trong khi bb khng c kh nng . Do khi gene tri C (khng c kh nng t ch) ng ring s cho mu nu; cn ng chung vi gene tri B s cho hiu qu b tr tri vi kiu hnh mu en. R rng cch gii thch ny ch hp l trn hnh thc khi cho rng allele tri C xc nh mu nu.
Hnh 2.8 T tri sang l cc chut agouti en, agouti nu v bch tng.
Kim chng: Ptc Chut nu (bbCC) Chut bch tng (BBcc) F1 Tt c chut en (BbCc) F1F1 = BbCc BbCc = (Bb Bb)(Cc Cc) F2 = (3B-:1bb)(3C-:1cc) = 9 B-C-: 3 bbC- : (3 B-cc + 1 bbcc) = 9 en : 3 nu : 4 bch tng 2.2. t ch do gene tri vi t l 12:3:1 V d: S di truyn mu sc lng nga. Khi lai gia hai ging nga
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thun chng lng xm v hung vi nhau, F1 thu c ton nga lng xm. Sau khi cho tp giao cc nga F1 th F2 c t l kiu hnh xp x 12 xm : 3 en : 1 hung . Gii thch: gii thch kt qu ny, ta quy c: A-B- v A-bb : xm; aaB-: en; v aabb: hung . Theo , allele tri B quy nh mu en l tri so vi b- hung . V s d A-B- v A-bb cho kiu hnh lng xm trong khi chng c cha B v bb, bi v gene tri A t ch cc gene khng allele v ng thi n cn c kh nng to mu xm, cn allele a khng c c hai kh nng . iu ny c th hin qua s lai sau y: Ptc Nga xm (AABB) Nga hung (aabb) F1 Tt c nga xm (AaBb) F1F1 = AaBb AaBb = (Aa Aa)(Bb Bb) F2 = (3A-: 1aa)(3B-:1bb) = (9 A-B- + 3 A-bb) : 3 aaB- : 1 aabb = 12 xm : 3 en : 1 hung 2.3. t ch do gene tri vi t l 13:3 V d: S di truyn mu sc lng g. Khi lai gia ging g thun chng, g Leghorn trng vi g Wyandotte trng, F1 thu c ton g lng trng. Sau khi cho tp giao cc c th F1, F2 c t l phn ly kiu hnh xp x 13 lng trng (white): 3 lng c mu (colored). Gii thch: gii thch kt qu ny, ta quy c: I-C- , I-cc v iicc : trng; v iiC-: c mu. Theo , allele tri C - gene sn xut cht to mu (chromogen) l tri so vi allele c- khng c kh nng to mu; v allele tri I (inhibitor) t ch gene khng allele vi n v n cng khng c kh nng to mu, cn allele i khng c kh nng t ch ln to mu. S lai: Ptc G Leghorn trng ( IICC ) G Wyandotte trng (iicc) F1 Tt c g trng (IiCc) F1F1 = IiCc IiCc = (Ii Ii)(Cc Cc) F2 = (3I-:1ii)(3C-:1cc) = (9 I-C- + 3 I-cc + 1 iicc) : 3 iiC= 13 trng : 3 c mu 3. Tng tc cng gp - s di truyn a gene v cc tnh trng s lng 3.1. Tng tc cng gp (additive) Tng tc cng gp hay s di truyn a gene (polygenic) l hin tng di truyn c trng ca mt s tnh trng s lng (quantitative trait), trong cc gene khng allele tc ng cng hng ln s biu hin ca mt tnh trng. Mi allele (thng l tri) ca cc gene a phn nh
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th ng gp mt phn ngang nhau trong s biu hin ra kiu hnh mt mc nht nh. Nh vy, liu lng cc allele tng dn trong cc kiu gene s to ra mt dy bin d kiu hnh lin tc trong qun th. V d: Cc th nghim ni ting nm 1909 ca nh di truyn hc Thy in (Sweden) Herman Nilsson-Ehle v s di truyn mu sc ht la m (ht y tc l phi nh - kernel). Khi lai gia cc ging la m thun chng ht vi ht trng, F1 ng thu c ton dng trung gian c mu hng; v ty theo dng ht c s dng trong cc th nghim m F2 s c cc t l phn ly gia ht c mu vi ht khng mu (trng) l 3:1, 15:1 hay 63:1. Kt qu phn tch cho thy chng do 2-3 gene a phn chi phi. Sau y ta hy xt trng hp F2 vi t l 15 c mu :1 khng mu, hay c th l 1 : 4 nht: 6 hng: 4 hng nht:1 trng. Gii thch: Do F2 c 16 kiu t hp vi t l tng ng trong khi F1 ng nht kiu gene, chng t F1 cho 4 loi giao t vi t l ngang nhau ngha l d hp t v hai cp gene phn ly c lp. y, F1 biu hin kiu hnh trung gian ca hai b m v F2 xut hin mt dy bin d lin tc cng hng. iu chng t tnh trng ny tun theo quy lut tc ng cng gp hay a phn tch ly. Quy c: V allele cho mu l tri hn allele cho mu trng v mc biu hin ca cc ht c mu F2 ty thuc vo liu lng cc allele trong kiu gene, nn ngi ta thng k hiu cc gene khng allele bng cc ch s 1,2...km theo cc ch ci in hoa (A) cho allele tri v ch ci in thng (a) cho allele ln, nh sau: A1, A2 - ; a1, a2 - trng. T y ta c th d dng xc nh kiu gen ca F1 (A1a1A2a2) v ca b m P: (A1A1A2A2) v trng (a1a1a2a2), v thit lp s lai hnh 2.9. Ptc A1A1A2A2 () a1a1a2a2 (trng) F1 A1a1A2a2 (hng) F2
Allele tri Kiu gen 4 1A1A1A2A2 3 2A1A1A2a2 2A1a1A2A2 nht 4/16 2 4A1a1A2a2 1A1A1a2a2 1a1a1A2A2 hng 6/16 1 2A1a1a2a2 2a1a1A2a2 hng nht 4/16 0 1a1a1a2a2
Kiu hnh T l
1/16
trng 1/16
Hnh 2.9 Mt php lai ca la m v trng do hai gene chi phi, cho thy mi tng quan gia t l ca cc kiu hnh F2 v s allele tri.
Mt v d c o khc l trng hp di truyn s dy ht trn bp ng (xem trong Di truyn hc i cng - Dubinin 1981, tr.135-145).
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Nhn xt: (1)Bng cch v mt th biu din mi quan h gia s allele tri c mt trong kiu gen (trn trc honh) v cc tn s kiu hnh (trn trc tung) F2, ta s thu c mt ng cong phn b chun c dng hnh chung, gi l phn phi Gauss. Trong kiu hnh trung gian hay cc kiu gene cha hai allele tri (tng ng vi tr trung bnh) chim t l cao nht, cn cc kiu hnh hoc kiu gene hai u mt tng ng vi cc ngng "cc oan" bao gi cng chim tn s thp nht (xem Hnh 2.10). cng l quy lut chung cho tt c cc tnh trng s lng. (2) i vi cc tnh trng di truyn theo kiu a phn cng gp, cc h s ca t l kiu hnh c th xc nh bng cch da vo tam gic Pascal. 3.2. Tnh trng s lng (quantitative trait) Thng thng, cc tnh trng c tr s kiu hnh lin quan n kch thc, trng lng hay hnh dng...c xc nh da trn thang nh lng (quantitative scale), c gi l cc tnh trng s lng.
(a)
(b)
Hnh 2.10 M hnh n gin v (a) s phn b chiu cao ngi (n v tnh y l inch, 1 in = 2,54 cm; trc tung ch s lng); v (b) tng quan gia cc mc mu da (trc honh) v cc t l trong qun th (trc tung).
Ni chung, cc tnh trng s lng c cc c im sau: Do nhiu gene quy nh; chu nh hng ln ca cc iu kin mi trng; v c s phn b kiu hnh lin tc trong mt qun th (hnh 2.10), nhng chng cng c th xy ra di dng cc lp kiu hnh khc nhau, chng hn nh trong cc v d v dy mu sc ht la m hoc s dy ht trn bp ng ni trn. V vy, i vi cc tnh trng ny, khng c mt mi quan h chnh xc gia tr s kiu hnh v mt kiu gene c th. Chng hn, ngi, l cc tnh trng v chiu cao, trng lng, hay ch s thng minh (intelligence quotient - IQ); cy la, la m l s ht trn mi bng,
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s bng trn mi khm...Tuy nhin, trong nhng nm gn y nh s dng cc ch th phn t (molecular marker), ngi ta tin hnh lp bn cc gene c hiu qu ln ln cc tnh trng c bit (nh cc bnh phc tp ngi, nng sut cy trng...) gi l QTLs, tc cc locus tnh trng s lng (quantitative trait loci). nghin cu s bin i kiu hnh ca cc tnh trng s lng, nht thit phi s dng cc phng php ca thng k ton hc. Chng hn, phng php ly mu sao cho hp l, ngha l mu phi ln, mang tnh ngu nhin v i din; vic x l s liu i hi phi s dng mt s i lng hay tham s thng k c bn sau y: - Trung bnh cng (mean):
x=
vx =
1 n xi n i =1
- Bin lng (variance):
1 n ( xi x) 2 n 1 i =1
- lch chun (standard deviation) bng cn bc hai ca bin lng (hay phng sai mu). Mt s i lng khc, nh h s bin thin (coefficient of variation, Cv%), h s tng quan (r) v.v... c th xem trong cc gio trnh Di truyn hc s lng v Thng k lin quan (Trn Vn Din v T Cm T 1995; Falconer 1983; Phan Hiu Hin 2001). 3.3. M hnh cc tnh trng s lng hiu v xc nh c tm quan trng ca cc tnh trng s lng, ta cn phi xy dng mt m hnh cho php chia ct cc tr s kiu hnh thnh ra cc thnh phn di truyn v mi trng. iu ny c th thc hin theo cch n gin bng cch biu th tr s kiu hnh (P: phenotype) cho mt kiu gene (i) trong mt mi trng c th (j) nh sau: Pij = Gi + Ei trong Gi l phn ng gp v mt di truyn ca kiu gene (genotype) i vo kiu hnh, v Ej l sai lch do mi trng (environment) j. Ej c th m hoc dng ty thuc vo s tc ng ca mi trng j. Cn lu rng, trong nhiu trng hp, cc qun th khc nhau c cc tr s trung bnh khc nhau; v tht kh m xc nh s khc nhau l do cc nhn t di truyn, nhn t mi trng, hay l s kt hp ca c hai gy ra. Cc kiu gene khc nhau c th tng tc mt cch khc nhau vi mi trng ca chng to ra kiu hnh. Nu nh cc mi tng tc c th nh th xy ra gia cc kiu gene v cc mi trng, khi ta c th m rng m hnh c s ni trn thnh m hnh tng tc kiu gene-mi
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trng (genotype-environment interaction), vi tr s kiu hnh l: Pij = Gi + Ej + GEij trong GEij l s o s tng tc gia kiu gene i v mi trng j. V mi trng bin ng nn s tng tc c th m hoc dng. Ni chung, m hnh di truyn c th c nh ngha mt cch chnh xc hn bng cch phn chia tr s kiu gene ra hai thnh phn nh sau: Gi = Ai + Di trong Ai v Di l cc thnh phn cng gp (additive) v tri (dominant). Nu nh th d hp ng l trung gian gia cc thnh phn ng hp (nh v d v s di truyn mu sc ht la m ni trn), th s ng gp ca thnh phn tri l zero; ngha l, ton b thnh phn di truyn l cng gp. Ni cch khc, nu cho rng s tng tc kiu gene-mi trng l khng ng k, th tr s kiu hnh c th c biu th bng: Pij = Ai + Di + Ej Nh ni t u, khng c s tng ng chnh xc gia mt kiu gene vi mt kiu hnh i vi cc tnh trng s lng. V vy, cch thc t hn kim tra cc tnh trng ny l tch s bin i trong mt qun th i vi mt kiu hnh c th thnh ra hai phn, phn bin i do nhn t di truyn gy ra v phn bin i do nhn t mi trng gy ra. Khi phng sai hay bin lng kiu hnh (phenotypic variance) s l: VP = VG + VE trong VP, VG v VE tng ng l cc bin lng ca kiu hnh, di truyn v mi trng. V theo cch lm trn, nu ta tch cc thnh phn cng gp v tri kim tra bin lng kiu hnh, ta c: VP = (VA + VD) + VE trong VA v VD tng ng l cc bin lng di truyn do cc hiu qu cng gp v tri. Nu ta chia biu thc ny cho VP, lc ta thu c t l ca bin lng kiu hnh do cc thnh phn khc nhau gy ra. C th l, t s gia bin lng di truyn cng gp v bin lng kiu hnh c coi l h s di truyn (h2; heritability): h2 = VA/VP i lng h s di truyn ny rt quan trng trong vic xc nh tc v hm lng p ng i vi s chn lc nh hng (directional selection), tc chn lc o thi cc kiu hnh cc oan, v n thng c cc nh chn ging ng vt v thc vt c tnh trc khi bt u mt chng trnh chn lc nhm ci thin nng sut hoc cc tnh trng khc. 3.3. c lng h s di truyn (heritability)
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By gi ta th tm hiu vn c lng h s di truyn da trn h s tng quan ca mt s tnh trng cc cp sinh i cng trng (monozygotic twins) v sinh i khc trng (dizygotic). Ta bit rng nhng cp sinh i ging ht nhau (cng trng) th c cng kiu gene; nh vy bt k s sai khc no gia chng phi l kt qu ca cc nhn t mi trng. Trn thc t, cc cp sinh i ging ht nhau l do s chia x cng kiu gene cng nh cc iu kin mi trng nh nhau. Mt cch xc nh mc tng ng hay ging nhau l tnh h s tng quan r (correlation) gia cc c th v cc tnh trng khc nhau. Tr s cao nht ca h s r l 1, trong tt c cc cp c th u c cng tr s kiu hnh nh nhau. Nu nh cc tnh trng l khng c tng quan gia cc cp c th th r = 0. Mt kt qu nghin cu v cc h s tng quan ca bn tnh trng khc nhau cc cp sinh i cng trng (rM) v cc cp sinh i khc trng (rD) nhng cng gii tnh c cho bng 2.3. Nhng sai khc gia cc tr s ny phi l kt qu ca s tng ng di truyn thp hn cc cp sinh i khc trng vn chia x (tnh bnh qun) ch mt na cc gen ca chng ni chung. Cng thc tnh h s di truyn nh sau: h2 = 2(rM rD) trong rM v rD ln lt l cc h s tng quan gia cc cp sinh i cng trng v khc trng. Bng cch s dng cng thc ny, ta tnh c cc h s di truyn cho bn tnh trng bng 2.3 bin thin t 0,98 n 0,16. Qua cho thy, hu ht s bin i cc ch s IQ v trng thnh v mt x hi dng nh l do mi trng, trong khi hu nh s bin i trong s lng np vn tay v chiu cao dng nh l do di truyn.
Bng 2.3 S tng quan ca bn tnh trng gia cc cp sinh i ngi Tnh trng S lng np vn tay Chiu cao im IQ im trng thnh x hi H s tng quan rD rM 0,96 0,47 0,90 0.57 0,83 0,66 0,97 0,89 H s di truyn 0,98 0,66 0,34 0,16
IV. Cc mi quan h kiu gene-kiu hnh

1. Thng bin v mc phn ng i vi cc tnh trng s lng ni trn, s biu hin ra kiu hnh ca mt kiu gene no r rng l ty thuc ca cc nhn t mi trng, tc l ty thuc ty thuc vo thm nhp v biu hin ca kiu gene (nh chng ta s xt di y). Hay ni cch khc, kiu hnh l kt qu ca s tng tc gia kiu gene v mi trng.
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Trong mt tnh hung l tng, phm vi tc dng ca mi trng ln mt kiu gene no c th xc nh c bng cch quan st kiu hnh ca cc c th ging nhau v mt di truyn c nui dng chm sc trong cc mi trng khc nhau. ngi, cc cp sinh i cng trng c tch nui ring lc s sinh chng minh cho mt tnh hung nh vy. Tuy nhin, nhng ch dn tt nht v cc tc dng ca mi trng bt ngun t cc nghin cu loi m c th nui trng vi s lng ln trn cc mi trng c kim sot. Th d kinh in cho nghin cu nh vy c J.Clausen v cc cng s tin hnh trong thp nin 1940. xc nh hiu qu ca mi trng ln cc kiu hnh, cc nh nghin cu ny trng cc dng (clones), tc gm cc c th ging nhau v mt di truyn, ct theo cc v khc nhau. Chng hn, h thu thp loi thuc h hoa hng l Potentilla glandulosa t ba vng: phm vi ven b bin Thi Bnh Dng v 100 mt; vng lng chng dy ni Sierras v 1.400 mt (ngn Mather); v cao 3.040 mt ca dy Sierras (ngn Timberline). T cc mu ny, h to ra c cc dng bng cch nhn ging sinh dng v em trng ba vng khc nhau . Ni khc i, mi kiu gene thu c t mt trong ba vng c a trng c ba iu kin mi trng (ng vi cc v ) khc nhau. T th nghim ny cho thy rng dng nh c hai nhn t di truyn v mi trng u quan trng trong vic xc nh s sinh trng ca cy. Th d, cc kiu gene vng ven b sinh trng tt c v thp v trung bnh nhng nhng khng th sng c v cao. iu ch ra tm quan trng ca cc nhn t di truyn. Cch thc m kiu hnh thuc mt kiu gene no thay i theo mi trng ca n c gi l mc phn ng (norm of reaction). Cc mc phn ng c th sai khc rt ln trong s cc kiu gene i vi cc tnh trng lin quan n tp tnh hay hnh vi (behaviour). D cho n nay ta khng h xem tp tnh hay hnh vi nh l mt kiu hnh, th nhng mt s cc gene t bin c nh hng ln tp tnh hay hnh vi. Chng hn, cc c th b bnh PKU khng c iu tr th c cc ch s IQ thp hn bnh thng; IQ l mt tnh trng c th o c bng trc nghim IQ. Tm li, mi quan h gia kiu gene v kiu hnh l phc tp, khng d g cn c vo kiu hnh xc nh kiu gene. Mc phn ng trn cc mi trng khc nhau l mt cch nh lng mi quan h . 2. thm nhp (penetrance) v biu hin (expressivity) nh gi mc th hin ra kiu hnh ca cc kiu gene nh th, nm 1925 Timofeev-Rissovsky nu ln hai khi nim: thm nhp v biu hin ca gene.
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2.1. thm nhp (penetrance) Trong cc th d xt trc y, nh by gene xc nh by tnh trng u H Lan, cc kiu nhm mu v nhiu bnh di truyn ngi u cho thy mt kiu tuyt i chc chn. Tuy nhin, i vi mt s gene, th mt kiu gene no c th hoc khng th cho thy mt kiu hnh no . Hin tng ny c gi l thm nhp (penetrance) ca mt gene. Mc thm nhp (level of penetrance) ny c th tnh bng t l ca cc c th mang mt kiu gene no bc l ra mt kiu hnh c th. Khi tt c cc c th ca mt kiu gene c th c cng kiu hnh nh nhau, th gene cho thy thm nhp hon ton v mc thm nhp ny c coi l bng 1. Mt khc, gene c coi l thm nhp khng han ton v mc thm nhp c th c th tnh c. Chng hn, trong s 160 c th thuc mt kiu gene no c 30 c th biu hin mt kiu hnh b bnh, th mc thm nhp l 30/160 = 0,1875; ngha l gene ch th hin ra c kiu hnh c 18,75%. Nh vy s c mt ca thm nhp khng hon ton ti mt gene c th khin cho mt kiu hnh tnh trng tri (gi s kiu gene l Aa) b ngt qung mt th h trong mt ph h, ri li xut hin tr li th h tip theo. l do allele tri thm nhp khng hon ton. Chng hn, dng tri ca nguyn bo li (retinoblastoma), mt bnh gy cc u c tnh mt tr em, ch thm nhp khong 90%. 2.2. biu hin (expressivity) Ngoi ra, mt kiu gen c th no bc l ra kiu hnh k vng, tc mc biu hin (level of expression) hay biu hin (expressivity), c th sai khc i. Chng hn, mc d mt gene gy ra mt bnh c th pht hin c hu ht cc c th mang mt kiu gene no , nhng mt s c th c th b nh hng nng hn cc c th khc. Bnh Huntington ngi l mt tnh trng cho thy biu hin bin thin sai khc theo tui pht bnh. Bnh ny ni chung xy ra tui bt u trung nn, vi tui trung bnh bt u pht bnh l trn 30 tui, nhng i khi c th xy ra rt sm tui cha ln 10 hoc mt ngi gi hn. Mt s ngi mang kiu gene bnh ny cht v nhng nguyn nhn khc trc khi bnh biu hin, do thm nhp khng hon ton cng nh biu hin bin thin i vi gene gy ra. 2.3. Cc nhn t mi trng v cc gene khc 2.3.1. Mi trng v cc gene gy cht c iu kin Vn t ra l ci g l nguyn nhn ca thm nhp khng hon ton v biu hin sai khc? Trong mt s trng hp c th, thng thf
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khng th xc nh c nguyn nhn, nhng c mi trng v cc gen khc cng c coi l c nh hng ln thm nhp v biu hin ca mt gene. V d, rui gim, cc allele ca mt s gene tc ng ln sc sng v c th gy cht nhit tng cao trn 28oC nhng li nh hng cht t hoc khng nh hng nhit thp hn. Cc gene nh th gi l cc gene gy cht c iu kin (conditional lethals); ni chung chng cho thy cc hiu qu gy cht trong cc tnh hung mi trng cc oan. Nhit cng c th nh hng mnh m ln kiu hnh ca cc thc vt. Chng hn, cy anh tho c hoa khi trng 24oC nhng li c hoa trng khi trng nhit trn 32oC. mo Xim (Thi Lan) v th Himalaya, mu sc b lng ca chng ti sm hn chn, tai v mi ti v thn nhit cc b phn xa nht ca c th ny t ra thp hn. Nu nh chng nhit
Hnh 2.11 Mu lng ca mo Xim ti sm cc vng chn, tai v mi.
m p hn, th s sinh trng lp lng mi ti cc im ny s cho mu sng nht (hnh 2.11). 2.3.2. Cc gene sa i (modifier or modifying genes) Ni chung, mc biu hin ca mt tnh trng gia cc c th c quan h h hng thng ging nhau hn l gia cc c th khng c quan h h hng, khi c hai nhm c th c nui dng chm sc trong nhng mi trng gn ging nh nhau. Cc gen c nh hng th cp ln mt tnh trng nh th gi l cc gene sa i (modifier genes) v chng c th gy nh hng ln kiu hnh mt cch ng k. Ni cch khc, cc gene sa i l nhng gene gy bin i cc gene khc trong khi chng biu hin ra kiu hnh.
Hnh 2.12 Thy tinh th bnh thng (tri) v bnh c thy tinh th (phi) do mt gen tri gy ra c tc ng ca gene sa i.
nhiu ng vt, allele pha long (dilute allele) vn lm gim cng hnh thnh sc t (nh t mu en sang mu xm chut nh) khi
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trng thi ng hp l mt v d v gene sa i. Mc ca hin tng khng ui cc mo Manx bin thin t khng c cc t sng ui cho n c mt t t sng ui dnh lin nhau, hnh nh l do nh hng ca cc gene sa i ln s biu hin ca th t bin tri ny. ngi, l trng hp mt gene tri gy c thy tinh th (hnh 2.12). Gene ny gy ra nhng mc suy yu th lc khc nhau ty thuc vo s c mt ca mt allele c th v mt gene sa i i cng. 2.3.3. Hin tng sao hnh (phenocopy) Mt hin tng lin quan gi l sao hnh (phenocopy), xy ra khi cc nhn t mi trng gy ra mt kiu hnh c th m thng thng c xc nh v mt di truyn. Trong trng hp ny, mt mi trng cc oan c th lm gin on s pht trin bnh thng bng mt kiu ta nh ca mt gene t bin. Mt trng hp thm thng xy ra cc nc Ty u vo cui thp nin 1950 v u thp nin 1960 khi mt s lng cc chu b sinh ra c t chi b ngn i mt cch kinh khng. Hin tng bt thng ny ging vi mt dng ri lon di truyn ln. Ch sau khi c mt cuc nghin cu rng ri ngi ta mi hay rng dng khuyt tt c gy ra v mt mi trng bi s b sung thalidomide vo bo ch vin thuc ng m cc b m ca cc chu b b bnh ung trong sut thi k u mang thai.
Cu hi v Bi tp
1. Hy ch ra cc mi quan h gia gene v tnh trng. Phn bit cc kiu tng tc gia cc gene khng allele ln s hnh thnh mt tnh trng. 2. c trng ca cc tnh trng s lng l g? Phn bit cc cp khi nim sau: tnh trng s lng v tnh trng cht lng, thm nhp v biu hin, tng tc gia cc gene allele v cc gene khng allele. 3. Mt ph n nhm mu M c mt a con mang nhm mu MN. B ta ni rng mt ngi n ng no c nhm mu MN l b ca n, v ng y tuyn b khng phi nh vy. (a) Vi thng tin , bn c th loi tr ngi n ng ny vi t cch ngi b khng? (b) Vic nghin cu xa hn cho thy ngi n b ny mang nhm mu B, a con mu O, v ngi n ng kia nhm mu AB. Vi thng tin ny, c th gt ngi n ng kia ra khi t cch ngi b hay khng? Ti sao? 4. c mo, cc php lai gia cc c th lng v bc i khi sinh ra tt c lng ; i khi : bc; v i lc l : trng: bc. Cc php lai gia hai con lng sinh ra hoc l ton b ; hoc : trng; hoc : bc. Hy xc nh phng thc di truyn ca tnh
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trng ny v kiu gene ca cc c th khc nhau tham gia vo cc php lai. 5. th, mu sc lng do bn allele ca mt gene n kim sot. Th t tri ca cc allele nh sau: C- xm > ca - sc len > cb - Himalaya > c bch tng. Ngoi ra, cc kiu gene cacb v cac u cho mu xm nht. Kt qu lai gia mt th xm vi ba con th khc nh sau: Php lai i con Xm sc len 6 xm, 5 xm nht Xm xm nht 8 xm, 3 xm nht, 4 himalaya Xm bch tng 9 xm, 8 himalaya Hy ch ra phng thc di truyn v kiu gene ca cc th b m. 6. Cc mu sc ca mt loi thc vt c th l , hng hoc trng. T kt qu di y, hy bin lun xc nh phng thc di truyn ca tnh trng mu hoa v ch ra cc kiu gene ca cc ging em lai. Php lai i con -1 Hng : hng -1 Trng : hng -2 Hng : hng : trng -3 Hng Tt c -3 Trng Tt c 7. Mu tc ngi gm c nm sc : vng, nu nht, nu va, nu thm v en. Cc php lai gia cc mu tc khc nhau cho kt qu sau: (1) vng vng tt c vng; (2) en en tt c en; (3) vng nu va tt c nu nht; hoc vng : nu va; (4) nu va nu va tt c nu va; hoc nu thm : nu nht; hoc nu va: en : vng. Hy xc nh kiu gene ca nm mu tc, v cho bit: (a) Nu lai gia nu nht v nu thm, t l k vng kiu hnh sinh ra nh th no? 8. chut, bt k c th no c cc l bch tng (albino). Hai cp allele khc, A/a v B/b, khi c mt allele C s cho mu xm, en, nu vng v nu sm. Bn dng thun chng bch tng khc nhau v kiu gene c cho giao phi vi cc con lng xm thun chng (AABBCC). Tt c chut cc i F1 u mu xm. Sau cc chut F1 t mi php lai c cho giao phi gia chng vi nhau, v kt qu cc i F2 nh sau y. Hy xc nh kiu gene mi dng bch tng.

Dng albino 1 2 3 4 Xm 44 31 48 145 en 0 0 15 43 Nu vng 16 0 0 46 Nu sm 0 0 0 15 Bch tng 20 9 21 82
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9. mt loi thc vt, khi lai gia mt dng hoa ta thun chng v ba dng hoa trng thun chng khc nhau vi kt qu cho nh sau:
Php lai 1 P F1 F2 ta trng-1 18 ta 95 ta 31 trng Php lai 2 ta trng-2 17 ta 80 ta 36 trng 28 Php lai 3 ta trng-3 19 ta 54 ta 32 trng 19 17 xanh da tri 6 nu
Hy bin lun v xc nh: (a) kiu gene ca bn ging hoa b m; (b) t l kiu hnh ca i con nu F1 ca php lai 2 c lai vi trng-2; (c) t l kiu hnh ca i con nu F1 t php lai 3 c cho lai vi trng-3. 10. Khi lai hai ging ng thun chng c 20 dy ht v 8 dy ht, tt c bp ng F1 u c 14 dy ht, v F2 c by kiu hnh khc nhau: 20, 18, 16, 14, 12, 10 v 8 (dy ht) vi s lng thng k tng ng l 40 : 245 : 594 : 793 : 603 : 236 : 38 (bp ng). Phng thc di truyn ca tnh trng ni trn l g? Gii thch, trnh by s lai v minh ho kt qu F2 bng mt th. Bn th rt ra quy lut chung v s di truyn ca loi tnh trng ny v ch ra cch xc nh t l kiu hnh k vng F2 i vi trng hp mt tnh trng c kim sot bi n locus c lp.
Ti liu Tham kho

Ting Vit Trn Vn Din v T Cm T. 1995. Di truyn s lng (Gio trnh cao hc Nng nghip). NXB Nng Nghip, H Ni. Dubinin NP. 1981. Di truyn hc i cng (bn dch ca Trn nh Min v Phan C Nhn). NXB Nng Nghip, H N. Hutt FB. 1964. Di truyn hc ng vt. (Bn dch ca Phan C Nhn). NXB Khoa hc v K thut, H Ni, 1978.
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Phan Hiu Hin. 2001. Phng php b tr th nghim v x l s liu (Thng k thc nghim). NXB Nng Nghip Tp.H Ch Minh. Phan C Nhn (ch bin), Nguyn Minh Cng, ng Hu Lanh. 1999. Di truyn hc. NXB Gio Dc, H Ni. Ting Anh Boon WH. 1984. Genes and the IQ. PG Publishing Pte Ltd, SingaporeHong Kong-New Dehli. Campbell NA, Reece JB. 2001. Essential Biology. Benjamin/Cummings, an imprint of Addison Wesley Longman, Inc, San Francisco, CA. Clegg CJ, Mackean DG. 2000. Advanced Biology: Principles and Applications. 2nd ed, John Murray Published Ltd, London. Falconer DS. 1983. Introduction to Quantitative Genetics. 2nd ed., Longman, London and New York. Hartl DL, Freifelder D, Snyder LA. 1988. Basic Genetics. Jones and Bartlett Publishers, Inc, Boston - Portola Valley. Suzuki DT, Griffiths AJF, Miller JH, Lewontin RC. 1989. An Introduction to Genetic Analysis. 4th ed, W-H Freeman and Company, New York. Weaver RF, Hedrick PW. 1997. Genetics. 3rd ed, McGraw-Hill Companies, Inc. Wm.C.Browm Publishers, Dubuque, IA. Wellnitz WR. 1995. Genetics: Problem Solving Guide. 2nd ed., Wm.C. Brown Publishers. Dubuque, Iowa. Mt s trang web http://www.biology.about.com/library/weekly/aa122602a.htm http://gslc.genetics.utah.edu/units/basis/ http://mhhe.com/lewisgenetics5 http://www.goodnet.com/nee72478/enable/hotline.htm http://www.cff.org http://consensus.nih.gov

Chng 3
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C s T bo ca s Sinh sn, Di truyn v Bin d

Nh chng ta u bit, cc khm ph ca Mendel v nhng nghin cu v cu trc t bo trong na cui th k XIX vn cn cha c s gn kt vi nhau. Vo nm 1902, Walter Sutton (USA) v Theodor Bovery (Germany), da trn tp tnh tng t ca cc gene v nhim sc th (chromosome) trong qu trnh gim phn gi rng cc gene c l nm trn cc nhim sc th. Sutton nhn nh: "Cui cng, ti xin lu rng xc sut kt hp ca cc nhim sc th b v m theo cc cp v phn ly sau trong s phn chia gim nhim... to thnh c s vt l cho cc quy lut di truyn Mendel". Ngy nay, iu y qu hin nhin. Nhng vo thi nhn nh ny ht sc quan trng; n to ra s kt ni cht ch gia di truyn hc v t bo hc, hnh thnh nn lnh vc mi gi l di truyn hc t bo vi s ra i ca thuyt di truyn nhim sc th. Trong chng ny, chng ta s tm hiu cc nhim sc th c hnh thi c trng nh th no; bng cch no s lng nhim sc th c duy tr n nh trong qu trnh nguyn phn, nhng li gim i mt na trong qu trnh gim phn; v cc kiu bin i v cu trc v s lng nhim sc th (t bin nhim sc th).
I. Sinh sn hu tnh v tnh n nh ca b nhim sc th

Trc tin, ta cp ch yu phng thc sinh sn hu tnh eukaryote v mi lin quan gia n vi s n nh v s lng nhim sc th c trng ca mi loi. Thc ra, cc eukaryote c hai kiu sinh sn chnh, v tnh v hu tnh. S sinh sn v tnh (asexual reproduction) xy ra khi mt c th n c to ra mt c th mi ging n; y l phng thc sinh sn ph bin thc vt v cc ng vt n gin. S trinh sinh (parthenogenesis) rp ci chng hn l mt trng hp c bit, cng sinh con nhng khng qua th tinh. Ni chung, con ci sinh ra bng cch ny th ging vi b m v mt di truyn. S sinh sn hu tnh (sexual reproduction) xy ra khi cc c th to ra cc t bo sinh dc c v ci, hay cc giao t (gametes), n lt chng kt hp vi nhau to thnh mt t bo trng c th tinh gi l hp t (zygote), tc mt t bo hon chnh m t pht trin thnh mt c th mi. Hnh thc sinh sn ny xy ra hu nh ton b cc kiu sinh vt, k
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c cc ng vt n gin nht nh con sum (Balanus) chng hn, cc thc vt, v thm ch c vi khun. vi khun, c cc kiu trao i thng tin di truyn nh tip hp, bin np v ti np c gi l sinh sn cn tnh (parasexual; vn ny c trnh by ring trong gio trnh Di truyn Vi sinh vt v ng dng). Thng thng th cc giao t c v ci bt ngun t cc c th khc nhau, cho nn i con sinh ra khc vi b m chng v nhiu chi tit. y l ni dung chnh m chng ny s tp trung tho lun. Cn s t th tinh c xem l trng hp ngoi l quan trng ca sinh sn hu tnh (xem chng 12).
Hp t ci (N)x Con ci trng thnh G Trng (2n) (2n) (n) Hp t (2n)
Hp t c (N)x Con c trng thnh G Tinh trng -------------[Sinh trng]------------------- * ----[Pht sinh]---- * --[Th tinh]-giao t / bo t
Hnh 3.1 S tng qut v sinh trng v sinh sn sinh vt hu tnh.

y cho thy s lng nhim sc th lng bi (2n) v n bi (n) tng ng vi cc giai on khc nhau (hng di cng). K hiu (N)x biu th nhiu ln nguyn phn, v G - gim phn.
S tng qut v s sinh trng v sinh sn ca sinh vt sinh sn hu tnh c trnh by hnh 3.1. Trn nguyn tc, mi hp t nhn c hai b nhim sc th n bi (haploid) k hiu l n, mt t giao t c v mt t giao t ci; nn s lng nhim sc th trong hp t l lng bi (diploid), tc 2n c trng v n nh cho loi. Mi b n bi cha n nhim sc th khc nhau, mi chic hay kiu nhim sc th ch c mt mt ln v cha cc gene khc nhau. Tp hp ton b cc gene trong mt b nhim sc th n bi nh th c gi l b gene (genome). Nh vy, trong b lng bi c trng ca cc t bo soma, cc nhim sc th tn ti theo tng cp gm hai chic ging nhau v hnh dng, kch thc v trt t phn b cc gene - mt c ngun gc t b v mt t m - gi l cc nhim sc th tng ng (homologous chromosomes). sinh vt a bo, hp t tng s lng t bo nh qu trnh nguyn phn (mitosis), l kiu phn chia t bo to ra cc t bo con c s lng nhim sc th 2n c gi nguyn. Khi c th t ti thnh thc sinh dc, mt s t bo ca c quan sinh sn tri qua gim phn (meiosis), tc kiu phn chia t bo to ra cc giao t c s lng nhim sc th gim i
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mt na (n). Qu trnh ny c gi l pht sinh giao t (gametogenesis) ng vt v pht sinh bo t (sporogenesis) thc vt. Sau , trong qu trnh th tinh (fertilization), cc giao t ci hay trng (egg) v giao t c hay tinh trng (sperm) hp nht vi nhau to thnh cc hp t i con. Cc hp t mi ny li bt u i vo mt chu k sinh trng v sinh sn y ht nh vy.
Bng 3.1 S lng nhim sc th 2n ca cc t bo soma mt s loi Loi ng vt 2n Loi thc vt Ngi (Homo sapiens) 46 u H Lan (Pisum sativum) Chimpanzee (Pan troglodites) 48 Ng (Zea mays) B (Bos taurus) 60 La go (Oryza sativa) Nga (Equus caballus) 64 La mch (Secal cereale) La (Equus asinus) 62 La m (Triticum durum) Ch (Canis familiaris) 78 La m (Triticum vulgare) Mo (Felis catus) 38 Khoai ty (Solanum tuberosum) Chut nht (Mus domesticus) 40 Thuc l (Nicotiana tabacum) G (Gallus domesticus) ~78 Loa kn (Lilium longiflorum) ch c (Rana pipiens) 26 Mn (Prunus domestica) C chp (Cyprinus carpio) 104 Bng (Gossypium hirsutum) Rui gim (D. melanogaster) 8 Hng dng (Helianthus annuus) 2n 14 20 24 14 28 42 48 48 12 48 52 34
Mi loi c mt s lng nhim sc th c trng; ngha l tt c cc c th ca cng mt loi th c s lng nhim sc th nh nhau. S lng nhim sc th ny hu ht cc loi ng-thc vt l lng bi (2n) c gii thiu bng 3.1, v mt s loi l n bi (n) - theo ngha c pha n bi l chnh - nh cc nm mc bnh m hng (Neurospora crassa, n = 7), mc bnh m en (Aspergillus nidulans, n = 8), nm men bia (Saccharomyces cerevisiae, n = 17)...R rng l s lng nhim sc th khng tng ng vi nc thang tin ha ca cc loi. Tuy nhin, ta c th thy cc loi c quan h h hng gn nhau nh ngi v chimpanzee hay nga v la th c s lng nhim sc th gn bng nhau. Cng cn lu rng, mt s loi thuc b cnh mng Hymenoptera nh ong mt (Apis mellifera) chng hn, con ci l 2n = 32 v con c l n = 16. i vi cc sinh vt ny ngi ta dng thut ng l n-lng bi (haplodiploid). mt s loi ng-thc vt nh ng, la mch en... bn cnh cc nhim sc th chun ca b nhim sc th bnh thng (gi l cc nhim sc th A), cn c thm mt vi nhim sc th rt b vn sai khc gia cc c th, gi l cc nhim sc th tha s (supernumerary chromosomes) hay nhim
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sc th B, phn bit vi cc nhim sc th A. Cc nhim sc th ny c coi l khng cha cc gen quan trng, mc d trong mt vi trng hp chng nh hng ln hu th. a s loi thuc lp chim (Aves), b nhim sc th cha cc nhim sc th rt nh gi l cc vi nhim sc th (microchromosomes) v thng c s lng ln.
II. Hnh thi hc nhim sc th eukaryote

tm hiu su hn v cu trc v chc nng ca cc nhim sc th, ta cn phi phn bit gia cc nhim sc th khc nhau v cc im sau y. 1. Kch thc nhim sc th Kch thc cc nhim sc th sai khc nhau rt ln. Gia cc loi khc nhau s sai khc ny c th ln ti hn 100 ln, trong khi mt s nhim sc th trong mt loi c kch thc hn km nhau khong 10 ln.
v tr tm ng chiu di kiu bng
(a) (b) Hnh 3.2 (a) B nhim sc th c x l bng thuc nhum ph bin Giemsa; v (b) xc nh cc c im ca mt nhim sc th.
m t b nhim sc th ca mt loi v xc nh cc nhim sc th ca n, cn phi thit lp kiu nhn (karyotype) v v s dng mt h thng cc quy c da trn kch thc ca chng, v tr tm ng, v cc kiu bng (cc hnh 3.2 v 3.5). Theo truyn thng, ngi ta s dng k thut hin bng (v d bng G hnh 3.2; xem thm mc 3) v chp nh cc nhim sc th k gia ca nguyn phn, gi l phng php k gia. Sau phng i nh, ct ri tng chic (gm hai chromatid ch em) v sp thnh tng cp tng ng (vai ngn quay ln trn v vai di hng xung di), ri xp theo th t nh dn v kch thc v nh s t ln nht n nh nht. Chng hn, i vi ngi (xem cc hnh 3.3 v 3.5): (i) cc nhim sc th khng phi gii tnh gi l nhim sc th thng (autosome) c nh s t 1 n 22 trn c s chiu di, cn cc nhim sc th gii tnh X v Y c tch ring. (ii) Vai ngn ca nhim sc th c biu th bng p v vai di bng q da trn v tr ca tm ng. (iii) mi vai trc tin c chia thnh cc vng v sau c xc nh bng cc bng. V d, k hiu 9q34 c ngha l vai di ca nhim sc th s 9,
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vng 3, v bng 4. y l ni khu tr ca gene xc nh h nhm mu ABO.
Hnh 3.3 B nhim sc th ngi c thit lp bng phng php mi kiu nhn ph (spectral karyotype). Ngun: Schrock v cs (1996).
Gn y, Schrock v cs (1996) gii thiu cc phng php thit lp kiu nhn mi s dng cc thuc nhum hunh quang c kh nng bm vo cc vng c th ca cc nhim sc th, gi l thit lp kiu nhn ph (spectral karyotyping; Hnh 3.3). Bng cch s dng mt lot cc vt d (probes) c th c hm lng cc thuc nhum khc bit, cc cp nhim sc th khc nhau s c cc c trng ph ring. Mt c im dc o ca cng ngh ny l s s dng mt nhiu k (interferometer) ging nh cc nh thin vn dng o quang ph pht ra t cc v sao. Cng ngh ny cho php pht hin cc bin i nh v mu sc (m mt thng chng ta khng th pht hin) bng chng trnh my tnh ri sau phn loi tng cp nhim sc th theo cc mu sc khc nhau. Vic sp xp cc nhim sc th theo tng cp rt n gin v cc cp tng ng th c cng mu nh nhau, v cc sai hnh hoc cc trao i cho c th nhn bit c mt cch d dng hn. 2. Tm ng v cc kiu nhim sc th Cc nhim sc th thng khc nhau v v tr tm ng (centromere), l ni m cc si thoi bm vo trong qu trnh phn bo (hnh 3.4). Thng th mi nhim sc th ch c mt eo tht ln cha mt tm ng gi l eo s cp (primary constriction). (Thut ng kinetochore dng ch cu trc protein bao quanh vng tm ng). Ni chung, c ba kiu
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nhim sc th chnh: nu tm ng nm gn chnh gia, gi l nhim sc th tm gia (metacentric); nu tm ng nm lch v mt u, gi l nhim sc th tm u (acrocentric); v nu tm ng gn st u mt, gi l nhim sc th tm mt (telocentric). Ni cch khc, tm ng chia nhim sc th thnh hai vai hay cnh (arms) di ngn khc nhau; chng hn, trong khi nhim sc th tm gia c hai vai gn nh bng nhau, th nhim sc th tm lch c mt vai ngn v mt vai di. Cc kt qu nghin cu cho thy chut nh tt c 40 cp nhim sc th u thuc kiu tm gia, trong khi b nhim sc th ngi gm hai kiu tm gia v tm u, khng c kiu tm mt (hnh 3.3). Ngoi eo s cp, trn mt s nhim sc th c th cn c th c eo th cp (secondary constriction), v nu eo ny gn u mt v xy ra s tht su s to nn mt v tinh (satellite) ca nhim sc th ( ngi, l cc nhim sc th 13, 14, 15, 21 v 22; xem hnh 3.5b). Eo th cp thng cha cc gene tng hp cc RNA ribosome, tch t tm thi v hnh thnh nn t chc gi l hch nhn (nucleolus); n c mt trong nhn (nucleus) mt s giai an khc nhau ca s phn bo. Hai u mt ca mi nhim sc th eukaryote c cu trc c bit gi l telomere (s c tho lun chng 5) v thng c trnh t DNA khc vi phn cn li ca nhim sc th.
B i
Cc chomatid khng ch em Cp tng ng
Cc chromatid ch em
(a) (b) Hnh 3.4 (a) Mt cp nhim sc th tng ng kiu tm u k gia. (b) Mi chic ly ra t nhn k gia gm hai chromatid ch em dnh nhau tm ng, gi l b i (dyad) v c cc u mt c trng gi l telomere.
3. Cc kiu bng nhim sc th (chromosomal bands) Ngi ta c th xc nh cc vng khc nhau trn cc nhim sc th bng cc k thut hin bng (band techniques) c x l cc ha cht khc nhau, nh cc bng G, Q v R (tng ng l Giemsa, quinacrine v reverse bands- bng o ngc) (xem Verma v Babu 1995). Thc ra, thut ng chromosome (= chromo: mu + soma: th) theo ngha en c
73
ngha l "th bt mu". Cc k thut s dng cc thuc nhum khc nhau xc nh cc vng ca nhim sc th no lm hin ra cc vch, di hay bng (bands) c m nht, sng ti hoc hin th bng cc mu khc nhau. Chng hn, cc bng sm hn thng xut hin gn tm ng hoc cc u mt (telomeres) ca cc nhim sc th, trong khi cc vng khc bt mu yu hn nn nht hn. Cc vng bt mu m gi l cht d nhim sc (heterchromatin) v cc vng bt mu nht gi l cht ng nhim sc (euchromatin). Ni chung, cc vng c trng ny c duy tr n nh trong cc t bo hay cc c th khc nhau ca mt loi v do chng t ra hu ch cho vic xc nh cc nhim sc th c th. Hnh 3.5 trnh by kiu nhn v biu nhim sc th (idogram) ngi.
(a) (b) Hnh 3.5 (a) Kiu nhn, v (b) biu nhim sc th ngi.
Ngun: Current Opinion in Genetics & Development (1998)
Cc vng euchromatin thng tri qua mt chu k ng v m xon u n, tng ng vi s hot ng ca hu ht cc gene khu tr trn . Cc vng heterochromatin gm hai kiu: n nh (constitutive) v khng n nh (facultative). Heterochromatin n nh, tc l phn c nh ca b gene v khng bin i c thnh euchromatin; chng c coi l bt hot v mt di truyn. Ngc li, heterochromatin khng n nh bao gm c euchromatin l nhng vng bt gi mu v c c (theo ngha ng xon cht) c trng ca euchromatin trong mt s giai on pht trin; hay ni cch khc, thc cht l cc vng euchromatin nhng ch bin i thnh heterochromatin trong k trung gian (interphase) mt s m. Trn thc t, ngi ta cn pht hin mt s dng nhim sc th c bit c kch thc khng l, nh: (i) cc nhim sc th kiu chi lng hay bn chi n (lampbrush chromosomes) bt gp trong non bo s cp ca
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ng vt c xng sng (c th di ti 1mm nh lng th c ui); v (ii) cc nhim sc th a si (polytene chromosomes) c mt trong nhn t bo tuyn nc bt ca u trng b hai cnh (Diptera).
(a) (b) Hnh 3.6 (a) Nhim sc th khng l trong mt nhn t bo truyn nc bt ca u trng D. melanogaster, v (b) cu trc chi tit ca nhim sc th X c tch lm hai na, mi na c 10 vng v mi vng bt u bng mt bng mu m.
Chng hn, rui gim D. melanogaster, cc nhim sc th a si ny sinh ra do hin tng ni nguyn phn (endomitosis), tc cc nhim sc th t nhn i lin tip c chc ln nhng t bo khng phn chia v hu nh trng thi m xon. V vy chng c th cha ti c ngn si chromatid dnh nhau trong mt b, vi chiu di hn100 ln so vi cc nhim sc th bnh thng (hnh 3.6). V xc nh c tt c l 102 vng, vi hn 5.000 bng. Ring mt mnh nhim sc th X (tc nhim sc th s 1) c 20 vng c nh s t 1 n 20, vi hn 1.000 bng. hnh 3.7b cho thy n l nhim sc th tm mt, c tm ng vng 20 nm st u mt bn phi ca na di, v gene gy ra mt trng nm gn u mt bn tri vng 3 thuc na trn.
III. Chu k t bo v nguyn phn

Nguyn phn (mitosis) l qu trnh phn chia t bo trong cc t bo con c to ra c s lng nhim sc th ging vi cc t bo b m. Kiu phn bo ny c trng cho cc t bo soma, k c cc t bo sinh dc (2n) pha sinh sn ca s pht sinh giao t cc ng-thc vt (mc IV.2), v xy ra theo cp s nhn vi cng bi bng 2, ngha l: t mt t bo ban u tri qua k ln nguyn phn lin tip s cho ra 2k t bo ging
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n. Nh vy m c th ln ln v cc t bo trong c th thng xuyn c i mi. Quy lut phn bo ny c minh ha n gin nh sau: 1 2 4 8 16 32 64 128 256 512 ... 1. Chu k t bo (cell cycle) Qu trnh nguyn phn lp li theo chu k nh vy c gi l chu k nguyn phn hay chu k t bo (cell cycle). Nguyn phn l mt phn ca ton b chu k t bo i vi cc t bo tri qua nguyn phn (nh hp t, cc t bo phi, cc t bo thuc cc m sinh trng hay m phn chia; hnh 3.7a). Ni chung, mt chu k t bo bao gm hai giai on chnh l nguyn phn (k hiu: M), l mt phn tng i nh ca ton b chu k
(a)
(b)
T bo ngng phn chia
Hnh 3.7 (a) S phn chia ca cc t bo chp r hnh ty (Allium cepa). (b) S tng qut ca mt chu k t bo.
t bo, v phn cn li ca chu k t bo gi l k trung gian (interphase). Gi l k trung gian bi v n nm gia hai ln phn chia lin tip. y l giai on t bo din ra cc hot ng chuyn ha cao , tng hp v ti bn vt cht di truyn - DNA, chun b tch cc cho t bo bc vo nguyn phn. N c chia thnh ba phn, gi l G1, S v G2. Nh vy, theo nguyn tc, mt chu k t bo bao gm bn giai on theo th t sau y (hnh 3.7b): (1) G1 (first gap) = giai an khi u trong t bo sinh trng, chuyn ha v chun b cho s ti bn b gene; (2) S (DNA synthesis) = tng hp DNA (v chi tit, xem chng 5); (3) G2 (second gap) = chun b cho qu trnh nguyn phn; v (4) M = nguyn phn (mitosis). Thi gian ca cc giai on khc nhau trong chu k t bo khc nhau mt cch ng k, ty thuc vo tng loi, tng kiu t bo, nhit v cc nhn t khc. Chng hn, thi lng tng ng vi bn giai on G1, S, G2 v M i vi cc t bo mu trng ca ngi ang phn chia l 11, 7, 4 v 2 gi (thi gian ton b l 24 gi). Khi mt hp t va c hnh thnh hay mt c th ang sinh trng,
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chu k ny c lp li nhiu ln hnh thnh nn mt c th vi hng t t bo. Mt s kiu t bo trng thnh, nh cc t bo thn kinh v t bo c vn gi nguyn k trung gian, thc hin cc chc nng c bit ha trong c th cho n lc cht v khng bao gi phn chia na; giai on c gi l pha G0. Tuy nhin, mt s t bo c th t pha G0 quay li i vo chu k t bo. Mc d hu ht cc t bo lympho trong mu ngi pha G0, nhng nu c s kch thch thch hp nh khi bt gp khng nguyn ph hp chng hn, chng c th b kch thch quay li chu k t bo. C th ni, G0 khng n thun ch ra s vng mt ca cc tn hiu cho nguyn phn m l mt s c ch hot tnh ca cc gene cn thit cho nguyn phn. Cc t bo ung th th khng th i vo pha G0 v c nh trc lp li chu k t bo mt cch v hn (xem chng 5).
K trung gian
u k trc
Cui k trc
K cui
K gia
K sau
Hnh 3.8 S biu din cc k ca nguyn phn v chu k ca n.
2. Nguyn phn (mitosis) Nguyn phn t n c th chia lm bn giai on khc nhau, din tin theo mt trnh t nh sau: k trc (prophase), k gia (metaphase), k sau (anaphase) v k cui (telophase). Mi giai on c mt nt c trng ring, c bit l mi lin quan vi tp tnh ca nhim sc th, nh m ta c th xc nh chng (hnh 3.8 v 3.9). Sau khi t nhn i k trung gian (c th l pha S) v hon tt vic chun b bc vo nguyn phn (pha G2), lc ny cc nhim sc th tip tc ng xon v kt c, nh vy chng hin r dn di knh hin vi quang hc. Mi nhim sc th by gi gm hai chromatid ch em (sister
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chromatids) dnh nhau tm ng. Theo nguyn tc, cc chromatid ny hon ton ging nhau do kt qu ca s ti bn bn bo ton DNA pha S (chng 5). Hn na, v hai chromatid ch em dnh nhau ti vng tm ng, nn chng c xem l mt nhim sc th.
Hnh 3.9 Cc giai on ca qu trnh nguyn phn mt t bo chp r hnh ty (Allium cepa).
2.1. K trc (prophase) Cng trong giai on ny, hch nhn (nucleolus) thng bin mt v mng nhn bt u tan v. Trung th (centriole) phn chia v hnh thnh xung quanh n mt cu trc mi gm rt nhiu si thoi (spindle fiber) tri di ti cc cc ca t bo. Mt s si thoi nh trc tip vo tm ng (c th l kinetochore) ca nhim sc th. 2.2. K gia (metaphase) Vo k gia, mng nhn tan bin hon ton, cc si thoi nh vo tm ng ca cc nhim sc th v y chng v mt phng xch o (equatorial plane) ca t bo v xp thnh mt vng. Ni chung, lc ny cc nhim sc th ng xon cc i (ngha l chiu di rt ngn ti a v do ng knh cng n ra ti a), vi cu trc in hnh c trng cho tng loi. Do , k gia l thi im thun li nht cho vic thit lp cc kiu nhn v nghin cu hnh thi hc cc nhim sc th nh ni trn. 2.3. K sau (anaphase) Vo u k sau, ti mi nhim sc th xy ra s phn tch tm ng, cc chromatid ch em by gi ri nhau v c gi l cc nhim sc th con (daughter chromosomes). K , cc si thoi co rt v gy ra s chuyn ng ca cc nhim sc th con ging nhau v hai cc i din. Nu nhn di knh hin vi lc ny, ta thy nhim sc th xut hin di
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dng ch V, J hoc I, ty theo kiu tm gia, tm u hay tm mt. Nh vy, chnh s sp xp thnh mt vng ca cc nhim sc th k gia v s phn ly ng u ca chng v hai cc k sau lm thnh bn cht hay l quy lut c trng cho qu trnh nguyn phn. 2.4. K cui (telophase) Vo k cui, hai b nhim sc th con v ti cc cc i din v bt u m xon. Lc ny mng nhn xut hin tr li v bao bc cc b nhim sc th; cc si thoi tan bin, hch nhn v cc nhn c hnh thnh tr li. K , ng vt, mng t bo hnh thnh mt eo tht (furrow) t ngoi vo trong; thc vt, mt phin t bo (cell plate) pht trin t trung tm ra ngoi. iu ny lm phn cch hai b nhim sc th con v t bo cht gia hai t bo con. Cc t bo con c s lng nhim sc th ging vi t bo ban u. Nh vy, thc ra, nguyn phn gm hai qu trnh phn chia: phn chia nhn (karyokinesis) v phn chia t bo cht (cytokinesis); nhng thc cht ca nguyn phn l s phn chia nhn.
IV. Gim phn, s pht sinh giao t v th tinh

1. Gim phn (meiosis) Gim phn l kiu phn bo c trng cho cc t bo sinh dc, trong cc t bo con sinh ra (gi chung l cc giao t) c s lng nhim sc th gim i mt na (hnh 3.10).
(3) gim phn II
(2) gim phn I
(1) k trung gian
cp nhim sc th tng ng
Hnh 3.10 Kt qu ca gim phn vi hai ln phn chia. y cho thy hu qu ca s ti t hp v phn chia gim nhim trong gim phn I.
Gim phn l mt giai on trong qu trnh pht sinh giao t, xy ra pha trng thnh (mature) sau khi b nhim sc th c nhn i k trung gian thuc pha sinh trng. Qu trnh gim phn gm hai ln phn
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chia ni tip nhau, gim phn I v gim phn II (hnh 3.10). Mi ln phn chia ny cng c chia lm bn k. Gim phn I (meiosis I) cn gi l phn chia gim nhim (reductional division), v s lng nhim sc th (2n) gim xung cn n bi (n). Trong gim phn I, cc chromatid ch em vn cn dnh nhau trong khi cc nhim sc th tng ng phn ly. Gim phn II (meiosis II) cn gi l phn chia ng u (equational division) v rt ging vi nguyn phn ch phn tch cc chromatid ch em v s lng nhim sc th gi nguyn khng i. Bi v gim phn l qu trnh di truyn quan trng v cn bn nht cp t bo, c s cho vic l gii cc quy lut di truyn v bin d, cho nn trc khi i vo m t gim phn, cn nm ba im chnh sau y: (1) Gim phn cng vi s th tinh sau cho php duy tr s lng nhim sc th cc loi sinh sn hu tnh. (2) Gim phn I cho php cc nhim sc th b v m khc nhau phn ly ngu nhin v mi giao t. (3) S trao i cho gia cc chromatid trn cc cp nhim sc th tng ng k trc trong gim phn I to ra cc t hp allele mi cc gene khc nhau. 1.1. Gim phn I (meiosis I) 1.1.1. K trc I (prophase I) y l pha phc tp nht ca ton b qu trnh gim phn, c chia lthnh nm giai on khc nhau (hnh 3.11a-e). Giai on th nht gi l giai on leptotene (si mnh), c trng bng s xut hin ca cc nhim sc th dng cc si mnh khi nhn di knh hin vi quang hc. K tip l giai on zygotene (si kt hp), cc nhim sc th tng ng tin li gn nhau v cc gene tng ng i din nhau. Qu trnh kt hp ca cc nhim sc th tng ng sau gi l tip hp (synapsis), l nt c trng c bn phn bit gia gim phn v nguyn phn. Cu trc gm hai nhim sc th tng ng cha bn chromatid kt cp nh vy gi l th lng tr (bivalent). Giai on th ba c tn l pachytene (si dy), v lc ny cc th lng tr ngn li v dy ln v s tip hp hon tt. Trong giai on ny c th xy ra hin tng trao i cho (crossing over) hay ti t hp (recombination) tng phn ca mi cp nhim sc th tng ng. Giai on th t gi l diplotene (si kp), cc nhim sc th tng ng bt u tch ra, c bit l cc vng nm hai bn tm ng. (D nhin, cc chromatid ch em cn dnh nhau tm ng cho ti u k sau II). Thng thng mi cp nhim sc th tng ng c th c mt hoc hai vng trong chng vn cn k st nhau hoc tip xc vi nhau, gi l cc hnh cho (chiasmata). Cc hnh cho ny (hnh 3.12) l bng chng
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vt l cho s ti t hp xy ra sau khi cc nhim sc th tng ng tip hp. Ni chung, c t nht mt hnh cho trn mt vai nhim sc th, nhng dc theo cc nhim sc th th c th c mt s hnh cho. Giai on cui ca k trc I, diakinesis, c c trng bng s ngn li ca cc nhim sc th v chm dt cc hnh cho. iu c ngha l, cc hnh cho b y v cc u mt ca cc nhim sc th. Lc ny, hch nhn v mng nhn cng bin mt.
Hnh 3.11 Cc giai on ca qu trnh gim phn hoa hnh ty (Allium cepa). K trc I gm 5 giai on: (a) leptotene, (b) zygotene, (c) pachytene, (d) diplotene, v (e) diakinesis. Tip theo l (f) k gia I, (g) k sau I, (h) k cui I, (i) k ngh ngn gia hai ln phn chia (interkinesis), (k) k gia II, (l) k sau II, v (m) k cui II.
1.1.2. K gia I (metaphase I) k gia I (hnh 3.11f), cc nhim sc th xp trn mt phng xch o thnh hai vng v cc tm ng c nh vo cc si thoi, sao cho c hai nhim sc th ca mi cp tng ng (tc th lng tr) nm i din nhau qua mt phng k gia, vi cc hnh cho xp thng hng dc theo n. S kin ny khc bit vi k gia nguyn phn v l c s cho s phn chia gim nhim v phn ly ngu nhin ca cc nhim sc th k sau I. 1.1.3. K sau I (anaphase I) Vo lc ny, hai tm ng trong cc nhim sc th tng ng ca
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mi th lng tr y nhau ra xa, sao cho mt nhim sc th ca mi cp tng ng i v mt cc (hnh 3.11g). Khi cc nhim sc th tng ng y nhau ra, cc hnh cho hon ton chm dt. Khc vi k sau nguyn phn, cc chromatid ch em k sau I vn cn dnh nhau tm ng.
(a)
(b)
Hnh 3.12 (a) Cc hnh cho trn mt cp nhim sc th. (b) S hnh thnh cc chromatid ti t hp bi mt hnh cho t hai nhim sc th tng ng.
1.1.4. K cui I (telophase I) hu ht sinh vt, sau khi cc nhim sc th di chuyn ti cc cc th mng nhn hnh thnh xung quanh chng v t bo ny phn chia thnh hai t bo con (hnh 3.11h). Tuy nhin, cc chi tit chnh xc v mt t bo hc ca k cui I cn nhiu sai bit, c bit l cc thc vt. 1.2. Gim phn II (meiosis II) Gia gim phn I v gim phn II thng ch c mt k trung gian ngn ngi gi l interkinesis (hnh 3.11i) Trong thi gian ny khng xy ra s tng hp DNA; mi t bo cha mt b nhim sc th n bi (n), trong mi nhim sc th cha hai chromatid ch em. K trc II (prophase II) thng xy ra rt nhanh v khng r nt; cc k cn li tng t nh trong nguyn phn. Tht vy, k gia II (metaphase II), cc tm ng nh vo cc si thoi v di chuyn v mt phng xch o (hnh 3.11k). Vo u k sau II (anaphase II), cc nhim sc th tch nhau tm ng v sau cc nhim sc th con phn ly v cc cc i din (hnh 3.11l). K cui II (telophase II) bt u khi ti mi cc c mt b nhim sc th n bi n (n) v mng nhn hnh thnh xung quanh chng (hnh 3.11m). Nh vy, theo nguyn tc, t mt t bo sinh dc 2n tri qua gim phn cho ra bn t bo sinh dc n bi, gi l cc giao t (hnh 3.10). 2. S pht sinh giao t (gametogenesis) Nh bit, gim phn l thi k hay pha (phase) quan trng nht ca qu trnh pht sinh giao t ng vt v pht sinh bo t thc vt. Di y ch cp ch yu qu trnh pht sinh giao t ng vt (hnh 3.13). ng vt, s to thnh cc giao t c hay tinh trng (sperm), gi l s sinh tinh (spermatogenesis), xy ra trong tinh hon (teste) - c quan
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sinh sn c. Qu trnh ny bt u vi s sinh trng ca mt t bo lng bi cha bit ha gi l t bo m tinh trng (spermatogonium). Ti pha sinh sn, t bo ny thc hin nhiu ln qu trnh nguyn phn gia tng s lng t bo m tinh trng (2n). Sau , mi t bo ny chuyn qua pha sinh trng, bit ha thnh mt tinh bo s cp (primary spermatocyte; 2n kp). K , t bo ny tri qua pha trng thnh vi hai ln phn chia lin tip ca gim phn. Sau gim phn I, mt tinh bo s cp cho ra hai tinh bo th cp n bi kp (secondary spermatocyte; n kp). Sau gim phn II, mi t bo ny phn chia thnh hai tinh t n bi (spermatid; n). Bc cui cng l s bit ha ca cc tinh t thnh cc t bo tinh trng (sperm), c cu to y cc b phn (nh u, c v ui di; hnh 3.13 v 3.14).
Hnh 3.13 S minh ha qu trnh sinh tinh v sinh trng ng vt.
S sinh trng (oogenesis) hay to cc giao t ci ng vt xy ra
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trong cc bung trng (ovary), c quan sinh sn ci. Qu trnh ny cng bt u bng mt t bo m ca trng (oogonium; 2n). Sau khi tri qua pha sinh sn, mi t bo sinh non ny bit ha thnh non bo s cp (primary oocyte; 2n kp) pha sinh trng, vi kch thc tng trng mt cch c bit. Ti pha trng thnh, sau gim phn I, t non bo s cp to ra mt non bo th cp (secondary oocyte; n kp) v th cc (polar body) th nht. Sau ln gim phn II, t non bo th cp cho ra mt non t (ootid; n) v mt th cc, cn th cc kia to ra hai th cc th hai. Chung cuc, t mt t bo sinh non (2n) cho ra ch mt non (n) kch thc rt ln v ba th cc (n) kch thc rt nh. Nguyn nhn l do gim phn xy ra gn mng t bo, nn cc th cc ch nhn c mt t t bo cht v chng vn cn bm trn b mt ca trng cho n lc tiu bin, tch ra. S tp trung t bo cht trong mt non t ri sau bit ha thnh trng (ovum, egg) chnh l ngun cung cp cht dinh dng cho s pht trin phi sau khi th tinh. S sinh tinh din ra lin tc cc ng vt trng thnh sinh sn quanh nm, chng hn nh ngi v ty theo ma cc ng vt sinh sn theo ma. Kh nng cho tinh trng l cc k cao hu ht cc ng vt; mt con c (male) trng thnh sn xut hng t t tinh trng trong sut qung i ca chng. Ngc li, vic sinh trng cc con ci (female) ni chung l thp hn rt nhiu; chng hn, mt ngi n sut i ch c th sn sinh chng 500 trng chn. Tt c s trng ny pht trin thnh cc non bo s cp trc khi con ci (female) c sinh ra v chng dng li k trc I cho ti tui dy th. Lc ny chng tip tc gim phn v bt u chn ln lt thnh cc trng v mi thng rng mt trng. T bo trng ngi c ng knh chng 0,1 mm (100 m) trong khi phn rng nht ca tinh trng c ng knh l 2,5 m. thc vt bc cao, s hnh thnh cc giao t c c gi l s hnh thnh ht phn hay pht sinh tiu bo t (microsporogenesis). Qu trnh ny ging nh s sinh tinh ng vt ch gim phn cho ra bn t bo n bi c cng kch thc v tim nng chc nng. S hnh thnh non thc vt cn gi l s hnh thnh ti phi hay pht sinh i bo t (megasporogenesis). N cng tng t nh qu trnh gim phn ng vt. Tuy nhin, cc sn phm sai khc nhau ng k. 3. S th tinh (fertilization) Ni chung, th tinh (hnh 3.14) l hin tng kt hp ngu nhin gia cc giao t c v giao t ci mang b nhim sc th n bi (n) to thnh cc hp t (zygotes) c b nhim sc th lng bi (2n) c trng ca loi. Thng thng, mt trng ch c th tinh bi mt tinh trng v to
84
ra mt hp t (hnh 3.14b). cc thc vt bc cao c qu trnh th tinh kp (double fertilization). Khi ng phn xm nhp ti phi (embryo sac) v a hai tinh t vo, s xy ra hai qu trnh th tinh: mt tinh t s kt hp vi non (egg) to thnh hp t (2n) v tinh t kia s dung hp vi nhn ni nh (endosperm nucleus; 2n) to thnh mt nhn ni nh (3n). Sau , trong qu trnh hnh thnh ht, hp t s pht trin thnh phi ht v nhn ni nh s pht trin thnh ni nh nui phi, gi l phi nh.
(a)
(b)
Hnh 3.14 (a) Cc tinh trng nhm bin (sea urchin) vy quanh mt trng nhm bin. (b) Mt t bo trng ngi va c th tinh, cc nhn ca tinh trng v trng th hin bng hai hnh dng khng u nhau, c mu vngnu. Khi hai nhn ny dung hp th s th tinh coi nh hon tt.
Ngun: (a) Francis Leroy, Biocosmos/SPL/Photo Researchers, Inc. (b) Phototake NYC/Dr. Nikas/Jason Burns 1993-2003 Microsoft Corporation.
V. Cc bin i ca nhim sc th
Trn y chng ta mi ch tm hiu cc c tnh di truyn n nh ca nhng c th mang b nhim sc th bnh thng m cha cp n mt bin i ca n. Trn thc t, cc nhim sc th c th b bin i (t bin) theo hai cch cn bn, l cc bin i v cu trc v v s lng (bng 3.2). Cc bin i ny c th gy nhng hu qu khc nhau ty loi, nhng thng l nghim trng. Chng c th gy hiu qu tc thi ln cc nhim sc th, gy ri lon trong hot ng ca b gene v gim phn; v t nh hng ti sc sng, hu th...ca cc c th mang chng. 1. Cc bin i v cu trc nhim sc th C bn kiu bin i cu trc nhim sc th, l: lp on (hay nhn on), mt an (hay khuyt an), o on v chuyn on nh gii thiu bng 3.2 (xem Hnh 3.15). Cc kiu t bin cu trc nhim sc th xy ra l do s t gy v ni li bt thng ca cc on trn cng mt cp tng ng hoc thm ch gia cc cp khc nhau nh trong trng hp chuyn on. Ni chung, cc t bin ny thng t gp, trung
85
bnh c 1.000 giao t mi hnh thnh c mt vi t bin lin quan kiu no . Cc hu qu v sau u l kt qu ca s tip hp gene-i-gene gia cc nhim sc th bnh thng v bt thng (kiu kt cp khng hon ton tng xng ny c gi l kiu d nhn, heterokaryotypic) cng nh cc vn sau ny xy ra cc k sau I v II trong gim phn.
Bng 3.2 Cc kiu bin i (t bin) nhim sc th Cc kiu bin i v cu trc Lp on (duplication) Mt on (deletion) o an (inversion) Chuyn on (translocation) Cc kiu bin i v s lng Mc nguyn bi (Euploidy) Cc th t a bi (autopolyploids) Cc th d a bi (allopolyloids) Lch bi (Aneuploidy) Cc th mt (monosomies; 2n 1) Cc th ba (trisomies; 2n + 1)
Mt on Lp on o on
1.1. Lp on (duplication) Lp on hay nhn on l trng hp mt on nhim sc th no c mt hai ln trn mt nhim sc th. Ty theo v tr v trt t ca cc vng lp an, ta c th phn ra hai kiu chnh sau: (i) Lp on c th nm k st vng nhim sc th ban u. Trong trng hp trt t cc bng (hoc cc gene) vn gi nguyn nh ban u th gi l lp an ni tip (tandem), hoc c trt t ngc li, gi l lp an o ngc (reverse). (ii) Nu nh vng lp li nm cch xa on gc ban u th gi l lp on chuyn ch (displaced). C th hnh dung cch lp on ni tip xy ra l do cc nhim sc th tng ng gi nhau v c cc ch t ng thi trn hai nhim sc th ti cc im khc nhau. Nu cc nhim sc th tng ng khc nhau ny ni li, th mt chic s c mt on lp ni tip v chic kia s mt i mt on tng ng.
Xen on
Chuyn on
Hnh 3.15 S tng qut cc kiu bin i cu trc nhim sc th.

Kiu xen on y l s chuyn an gia (interstitial translocation).
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Trong trng hp , lp on v mt on l hai kiu thun nghch ca nhau. iu cng c th do c ch trao i cho khng u (unequal crossing over) gy ra (Hnh 3.16).
Hnh 3.16 Lp on v mt on do c ch trao i cho khng u gy ra.
Hu qu l: (1) V phng din t bo hc, khi mt c th l d hp kiu nhn v mt nhim sc th lp on v mt nhim sc th bnh thng th vng lp on khng c on tng ng kt cp trong gim phn. Lc on lp s phnh ra di dng mt cu trc hnh vng (loop) sao cho cc phn cn li ca c hai nhim sc th c th bt cp vi nhau, tc tip hp gene-i-gene (Hnh 3.17). mt s trng hp, mt vai nhim sc th c cha on lp c th un vng li sao cho hai u ca on lp giao nhau.
Hnh 3.17 S kt cp ca cc nhim sc th cc c th c kiu nhn d hp v (a) lp on ni tip - tandem v (b) lp on o ngc - reverse.
(2) V phng din kiu hnh, cc c th d hp hoc ng hp v cc on lp b vn c th sng, mc d chng thng bc l ra mt vi hiu qu kiu hnh (nh trong trng hp cc t bin mt Bar rui gim). (3) V phng din tin ha, nu nh cc c th mang cc t bin ny vn sng bnh thng, th chng s c tim nng cho s bin i tin ha xa hn na cc gene ph thm ny, gi l tin ha bng c ch nhn i gene. Thc ra, c rt nhiu bng chng chng t s nhn i gene l mt
87
c ch quan trng nht sinh ra cc gen mi v cc qu trnh tin ha mi, to thun li cho s tin ha ca cc sinh vt phc tp t cc sinh vt nguyn s. Chng hn, cc gene khc nhau m ha cho hemoglobin cc ng vt c xng sng c th bt ngun t mt gene t tin c nhn i, v sau cc bn sao nhn i ny phn ly v mt chc nng ca chng (c th xem thm trong: Kimura 1983; Li 1983). 1.2. Mt on (deletion) Mt nhim sc th b khuyt mt i mt an c gi l t bin mt an (deletion) hay khuyt on (deficiency) (hnh 3.18). Mt on c th xy ra ti mt phn bn trong nhim sc th, gi l mt on gia hay mt an khe (interstitial deletion). Cn nu nh mt an nhim sc th b t v khng ni li c, gi l mt on mt (terminal deletion). Trong trng hp on t gy c cha tm ng th c on ny ln nhim sc th mt an thng s b dung gii trong qu trnh phn bo. C ch mt on do s trao i cho khng u ni ti phn trn.
(a)
(b)
Hnh 3.18 (a) C ch gy mt on mt (tri) v on khe; (b) Hi chng cri du chat do mt on mt trn vai ngn nhim sc th s 5.
Hu qu: Khi cc th mt an trng thi ng hp, chng thng b cht do khuyt hn cc gene thit yu. Thm ch khi trng thi d hp, cc gene gy cht c th gy ra s pht trin bt thng. Mt v d ni ting ngi l t bin mt on mt phn ng k ca vai ngn trn nhim sc th s 5 (5p), khi trng thi d hp gy ra hi chng "mo ku" (ting Php: cri du chat; ting Anh: cry-of-the-cat). Nhng a tr mc hi chng ny ni chung c ging cao the th c trng ging nh ting mo ku, c u nh v tr n; v chng thng b cht sm tui s sinh hoc th u (Hnh 3.18). Ngoi ra, cc th d hp v mt an thng cho thy s kt cp nhim sc th bt thng trong gim phn. Do n khng c vng tng ng m kt cp vi, nn trn nhim sc th bnh thng s hnh thnh mt ci vng gi l vng mt on (deletion loop). Tuy nhin, mt s c im khc li t ra hu ch cho vic xc nh cc mt an, chng hn: (1) Khng nh cc t bin khc, cc t bin
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mt an l khng o ngc tr thnh nhim sc th kiu di c (tc dng bnh thng vn c ca t nhin). (2) cc th d hp mt on, cc allele ln trn nhim sc th bnh thng s biu hin ra kiu hnh bi v nhim sc th mt on b khuyt vng tng ng. S biu hin ca cc allele ln trong trng hp nh vy, gi l hin tng tri gi (pseudodominance); n c ch trong vic xc nh chiu di ca on b mt. 1.3. o on (inversion) o an l hin tng xy ra do s t ng thi ti hai im trn mt nhim sc th v sau on b t xoay 180o ri ni li. Hu qu l, trt t cc gene trong on o ngc li vi trt t bnh thng (hnh 3.15). Ty theo s tng quan ca on o vi v tr tm ng, c th chia lm hai kiu o an. Nu an o khng cha tm ng, gi l o on cn tm (paracentric inversion); ngc li, nu on o bng qua c tm ng th gi l o on quanh tm (pericentric inversion).
Hnh 3.19 S trao i cho xy ra bn trong vng ca th d hp o on cn tm to ra cc giao t cha cc khuyt an ln. (a) Kt cp v xut hin vng; (b) Phn ly lm xut hin cu ni v on khng tm; (c) Cu ni hai tm t gy ngu nhin; v (d) Cc sn phm c to ra.
89
V c ch, c th hnh dung nh sau: v l do no , mt nhim sc th xon li mt vai (khng lin quan tm ng) hoc bng qua c hai vai (tm ng nm trn vng cun), sau xy ra s t gy ti vng giao nhau ri ni li khng ng. Cc c th d hp v mt on o c th nhn din bi s c mt ca cc vng o on (inversion loops) trn cc nhim sc th thuc giai an si dy (pachytene) ca gim phn. l do i lc ca hai nhim sc th tng ng. Cch duy nht m hai nhim sc th tng ng ny cp c vi nhau l mt chic t n xon li thnh mt cu trc hnh vng trong khi chic kia lm thnh mt na vng bao quanh n m khng xon li. Nu nh s trao i cho xy ra trong vng c on o (Hnh 3.19), c th dn ti hnh thnh cc giao t cha cc on lp v khuyt an ln v thng gy cht giai an hp t. 1.4. Chuyn on (translocation) Chuyn an l di chuyn ca mt on nhim sc th t mt nhim sc th ny sang mt nhim sc th khc, khng tng ng vi n v gy ra s cu trc li nhim sc th ng k (hnh 3.15, 3.20). C hai kiu chuyn an: (i) chuyn on gia (interstitial translocation) l s di chuyn mt on cho mt nhim sc th khc m khng nhn li, tc chuyn on khng thun nghch; v (ii) chuyn on thun nghch (reciprocal translocation), tc c s trao i qua li hai chiu. Kiu th hai ph bin hn. Nu nh hai on c ni li trong chuyn an thun nghch to ra hai chic c kch thc ln v hai chic kia b, th cc nhim sc th c chuyn on c kch thc b hn thng b bin mt. Khi , s lng nhim sc th b gim i do s trao i nhim sc th. Nh th, cc chuyn an c th lm thay i c kch thc ln v tr tm ng. Nhng c th d hp v chuyn on ch sinh ra khong mt na s con bnh thng; hin tng c gi l bn bt dc (semi-sterility). Khi cc th d hp chuyn on tri qua gim phn s cho ra khong mt na s giao t cha c hai phn ca chuyn on thun nghch v khong mt na kia cha c hai nhim sc th bnh thng. S giao t cn li ch cha mt phn ca chuyn an v mt nhim sc th bnh thng; cc giao t loi ny s gy cht hp t do mt cn bng v vt cht di truyn. in hnh l trng hp chuyn an Robertson (Robertsonian translocation), trong cc vai di ca hai nhim sc th tm u khng tng ng gn vi nhau v ch c mt tm ng (hnh 3.20b). Cn nhim sc th tm u b t gy vai di v on t t nhim sc th kia c ni li thng bin mt trong qu trnh phn bo hoc sau vi th h. Cc chuyn an Robertson c ngha quan trng trong di truyn hc ngi.
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(a)
(b)

NST 14
+
NST 21
NST (14 + 21q)
Hnh 3.20 (a) Chuyn on thun nghch; (b) Chuyn on Robertson, v s bn di cho thy s t - ni ca hai nhim sc th khng tng ng to ra kiu chuyn on ny. Mi tn () ch cc im t.
Mc d cc chuyn an c th to ra cc nhim sc th bnh thng, song chng cng c th gy ra nhiu bnh di truyn ngi. Chng hn, khong 5% s ngi mc hi chng Down (Down syndrome) c mt b m l d hp v mt chuyn an. C th trong trng hp ny, vai di nhim sc th 21 (an 21q) b t ra v gn vo ch t trn vai ngn nhim sc th 14; ngi b hoc m ny trng thi d hp vi kiu hnh bnh thng, nhng c nguy c gy hi chng Down i con do s phn ly sai hnh xy ra trong gim phn. Theo nguyn tc, khi gim phn, th d hp chuyn on c th to ra bn loi giao t, vi xc sut ngang nhau, thuc hai nhm c b nhim sc th cn bng v khng cn bng sau y: [14 ; 21] : [14 + 21q] : [14] : [(14 + 21q) ; 21] Khi cc giao t ny th tinh vi cc giao t bnh thng [14 ; 21] s cho ra bn kiu hp t tng ng l:
[(14)2 ; (21)2] : [(14;14 + 21q) + 21] : [(14)2 + 21] : [(14;14 + 21q) + (21)2]
Bnh thng : Th d hp chuyn an : Th mt 21 : Th ba 21
(46) (45) (45; cht) (46) Bi v cc c th nhn c ch mt nhim sc th 21 s b cht, nn chung cuc c khong 1/3 s tr sng (t mt th d hp chuyn on nh vy) c k vng l mc hi chng Down. Trn thc t, t l ny thp hn 1/3, ch yu l do mt s c th khng sng st c thi k thai. Cn lu rng, nguyn nhn ny ca hi chng Down a ra cc gi cho t vn di truyn (genetic counseling) ch: (1) Hi chng Down c th ti din cc con ci ca mt kiu nhn d hp v chuyn on, trong
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khi hi chng Down bnh thng th khng ti din trong cc anh ch em rut v v sinh (xem mc 2.2 di y). (2) Mt na s anh ch em rut bnh thng v kiu hnh ca cc c th Down chnh h li l nhng th d hp v chuyn an, v do c th sinh ra con ci b Down. nc ta, theo Nguyn Vn Rc (2004), c khong 6% trng hp hi chng Down do cc th chuyn on gy ra, bao gm cc dng t(14q; 21q), t(21q; 21q) v t(21q; 22q). Trn hnh 3.21 trnh by c ch hnh thnh cc loi giao t t ngi n thuc dng chuyn on t(21q; 22q) v s th tinh gia mt trng bt thng c nhim sc th 22 c chuyn on 21q (k hiu l: 23,X,+21q) vi tinh trng bnh thng (23,X) to ra hi chng Down thuc dng chuyn on (46,XX, +21q).
(a) (b) Hnh 3.21 Hi chng Down do th chuyn on dng t(21q; 22q) gy ra: (a) S hnh thnh cc loi giao t khc nhau t ngi n t(21q; 22q); v (b) S th tinh gia mt trng bt thng (23,X,+21q) vi tinh trng bnh thng (23,X) to ra hi chng Down chuyn on dng (46,XX, +21q).
2. Cc bin i v s lng nhim sc th T bng 3.2 cho thy s bin i s lng nhim sc th sai khc nhau hai cch c bn: (1) Cc th bin d nguyn bi hay th a bi (euploid variants) c s lng cc b nhim sc th khc nhau; v (2) cc th bin d lch bi hay th d bi (aneuploid variants) c s lng nhim sc th c th khng phi l lng bi. Ni chung, cc kiu bin i s lng nhim sc th nh hng ln s sng st ln hn cc kiu bin i cu trc nhim sc th. Trn thc t, ngi, hn mt na cc trng hp sy thai t nhin xy ra trong ba thng mang thai u c lin quan ti cc thai b lch bi, a bi, hoc cc sai hnh nhim sc th ln khc. Trc khi tho lun cc th t bin s lng nhim sc th lin quan ti s tng hoc gim hm lng vt cht di truyn, ta hy xt qua cc c ch khc m qua s lng nhim sc th c th thay i (hnh 3.22).

(a)
NST 1 (b)
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+
NST 2
NST ln (1q + 2q)
+
NST b
(c)
Hnh 3.22 S hnh thnh nhim sc th tm gia do dung hp tm ng ca hai nhim sc th tm u khng tng ng (a), v cc dng nhim sc th hai tm v khng tm (b v c).
Nh cp, nu nh cc chuyn an thun nghch xy ra gia hai nhim sc th tm u m cc an ln (tc vai di) c ni li vi nhau, kt qa l to ra mt nhim sc th ln c tm gia v mt nhim sc th b m n c th bin mt trong qu trnh phn bo. C ch hnh thnh mt nhim sc th tm gia ln do s t-ni xy ra ti cc tm ng ca hai nhim sc th tm u khng tng ng nh th gi l s dung hp tm ng (centric fusion; hnh 3.22a). Ngoi ra, thnh thong cng bt gp cc dng nhim sc th bt thng c hai tm ng v khng tm ng (hnh 3.22b-c). Tuy nhin, cc dng bt thng nh th c tnh khng n nh bi v chng thng b loi b khi t bo khi phn ly v hai cc trong qu trnh phn bo; chng hn, dng hai tm ng thng xut hin dng cu ni v s b t gy do cc si thoi ko cng v hai cc. 2.1. Hin tng a bi (polyploidy) Cc th a bi (polyploids) l trng hp trtong cc sinh vt c ba, b nhim sc th tr ln (hnh 3.23). Nu ta gi x l s nhim sc th n bi c bn, khi cc sinh vt c ba, bn b nhim sc th... s c s nhim sc th v tn gi tng ng l 3x (th tam bi: triploid), 4x (th t bi: tetraploid)... Lu : thay v dng k hiu n ch s nhim sc th n bi nh trc y, y x ch s nhim sc th trong mt b v n biu th cho s nhim sc th trong mt giao t. V d: mt sinh vt lc bi vi 60 nhim sc th (6x = 2n = 60), th x = 10 v n = 30.
Hnh 3.23 Hin tng a bi ha in hnh chi Chrysanthemum, vi rt nhiu loi a bi th khc nhau sinh ra t loi lng bi (2n = 18).
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Ni chung, hin tng a bi th tng i ph bin cc thc vt nhng him gp hu ht cc ng vt. Gn nh mt na s thc vt c hoa u l cc th a bi, k c cc loi cy trng quan trng. Chng hn, khoai ty t bi (4x = 48), la m mm lc bi (6x = 42), v cy du ty bt bi (8x = 56). thc vt bc cao, Chrysanthemum l mt chi in hnh v hin tng a bi ha (hnh 3.23). Trong qu trnh gim phn cc loi thuc chi ny, cc nhim sc th kt i to thnh cc th lng tr, loi 118 nhim sc th to thnh 9 th lng tr, loi 36 nhim sc th to thnh 18 th lng tr v.v... Mi giao t nhn mt nhim sc th t mi th lng tr, v vy s lng nhim sc th trong mi giao t ca bt k loi no cng chnh bng mt na s lng nhim sc th c tm thy trong mi t bo soma ca n. V d, loi thp bi c 90 nhim sc th th to thnh 45 th lng tr, do mi giao t s mang 45 nhim sc th. Nh vy qua th tinh, b y 90 nhim sc th ca loi ny c phc hi. Nh vy, cc giao t ca c th a bi r rng l khng phi n bi nh c th lng bi. Thng thng, ngi ta phn bit hai kiu th a bi: (1) cc th a bi cng ngun hay th t a bi (autopolyploids) l cc th a bi nhn c tt c cc b nhim sc th ca chng t cng mt loi; v (2) cc th a bi khc ngun hay th d a bi (allopolyploids) l cc th a bi nhn c cc b nhim sc th t cc loi khc nhau. Chng hn, nu nh mt ht phn lng bi khng gim nhim t mt loi lng bi th tinh cho mt trng lng bi cng ca loi , i con sinh ra l cc th t t bi (autotetraploids), hay AAAA, trong A biu th mt b nhim sc th hon chnh hay b gene (genome) ca kiu A. Mt khc, nu nh ht phn lng bi ca mt loi th tinh cho mt trng lng bi ca mt loi khc c quan h h hng vi loi ny, i con sinh ra s l cc th d t bi (allotetraploids), hay AABB, trong B ch b gene ca loi th hai. Tt c cc b nhim sc th trong mt th t a bi u l tng ng, ging nh khi chng trong mt th lng bi. Nhng trong cc th d a bi, cc b nhim sc th khc nhau ni chung sai khc nhau mt mc no , v c gi l tng ng mt phn (homeologous), hay tng ng tng phn (partially homologous). Trong t nhin, cc th a bi xy ra vi tn s rt thp, khi mt t bo tri qua s nguyn phn hoc gim phn bt thng. Chng hn, nu trong nguyn phn tt c cc nhim sc th i v mt cc, th t bo s c s nhim sc th l t t bi. Nu nh xy ra gim phn bt thng, c th to ra mt giao t khng gim nhim c 2n nhim sc th. Tuy nhin, trong hu ht cc tnh hung, giao t lng bi ny s kt hp vi mt giao t n bi bnh thng v sinh ra mt th tam bi. Ngi ta cng c
94
th to ra cc th a bi bng cch x l colchicine, mt loi ha cht gy ri lon s hnh thnh thoi v sc. Kt qu l, cc nhim sc th khng phn ly v cc cc c, v thng th xut hin cc th t t bi. 2.1.1. Cc th t a bi (autopolyploids) Cc c th tam bi (AAA) thng l cc th t a bi sinh ra do th tinh gia cc giao t n bi v lng bi. Chng thng bt dc bi v xc sut sinh ra cc giao t c c cn bng l rt thp. Trong gim phn, ba ci tng ng c th kt cp v hnh thnh mt th lng tr, hoc hai ci tng ng kt cp nh l mt th lng tr, li nhim sc th th ba khng kt cp. Tuy nhin, do tp tnh ca cc nhim sc th khng tng ng l c lp, nn xc sut mt giao t c chnh xc n nhim sc th l ()n (s dng quy tc nhn), v xc sut mt giao t c c chnh xc 2n nhim sc th cng l ()n. Tt c cc giao t khc cn li s l khng c s cn bng v ni chung l khng hot ng chc nng trong cc hp t c cha chng. Chng hn, hu ht cc cy chui l cc th tam bi; chng sinh ra cc giao t khng cn bng, v kt qu l khng c ht.
Qu bnh thng
Triticum monococcum (2n = 14)
Triticum kiu di (2n = 14)
Qu ca th a bi
Con lai bt dc Sai st do gim phn
T. turgidum (2n = 28)
T. fauschii (kiu di) (2n = 14) Con lai bt dc
Hnh 3.24 S hnh thnh th a bi vi hoa tri ln hn th 2n bnh thng. Hnh 3.25 S hnh thnh la m Triticum aestivum d lc bi (2n = 42) bng con ng d a bi.
Sai st do gim phn
T. aestivum (2n = 42)
Cc th a bi thng ln hn cc th lng bi h hng (v d, cho hoa tri ln hn; Hnh 3.24). Cc th t a bi c th gim phn bnh
95
thng nu nh chng ch to thnh cc th lng tr hoc cc th t tr. Cn nu nh bn nhim sc th tng ng to thnh mt th tam tr v mt th n tr, th cc giao t ni chung s c qu nhiu hoc qu t cc nhim sc th. 2.1.2. Cc th d a bi (allopolyploids) Hu ht cc th a bi trong t nhin u l cc th d a bi, v chng c th cho ra mt loi mi. Chng hn, la m Triticum aestivum l mt dng d lc bi vi 42 nhim sc th. Qua kim tra cc loi hoang di h hng cho thy la m ny bt ngun t ba dng t tin lng bi khc nhau, mi dng ng gp hai b nhim sc th ( y ta k hiu AABBDD). S kt cp ch xy ra gia cc b nhim sc th tng ng, v vy gim phn l bnh thng v cho ra cc giao t cn bng c n = 21. R rng, hin tng d a bi ng vai tr quan trng trong s tin ho ca la m (xem Hnh 3.25). Nm 1928, nh khoa hc ngi Nga, G. Karpechenko to ra mt th d t bi rt l c bit; khi ng lai gia ci bp Brassica v ci c Raphanus sativus, c hai u c s nhim sc th lng bi l 18. ng mun to ra con lai c l ca cy ci bp v c ca cy ci c. Sau khi thu c cc ht lai t mt cy lai nhn to, em gieo trng v pht hin rng chng c 36 nhim sc th. Tuy nhin, thay v thu c cc tnh trng nh ng mong i, cy lai ny c l ca cy ci c v c ca cy ci bp! Hnh 3.26 cho thy s hnh thnh con lai gia hai loi ci c v ci bp ni trn, c gi l Raphanobrassica.
Giao t
B m
Con lai F1 bt th Th song nh bi hu th
Hnh 3.26 S to thnh th song nh bi hu th Raphanobrassica t hai loi ci bp Brassica v ci c Raphanus u c 2n = 18 (tri); v mt kt qu c th ca con lai F1 Raphanobrassica (theo W.P.Amstrong 2000).
96
Trong trng hp nu mt ht phn n bi c b gene A th phn cho hoa ca loi c b gene B, s cho ra mt con lai bt th c thnh phn b gene AB. Nu nh sau nguyn phn khng xy ra c trn mt nhnh, c th sinh ra cc t bo AABB. Nu cc t bo ny t th phn th s to ra mt th d a bi. Li dng c im ny, cc nh chn ging s dng colchicine tc ng ln con lai bt th to ra cc th d a bi. 2.2. Hin tng lch bi (aneuploidy) Trong t nhin, thnh thong ta bt gp cc c th c s lng nhim sc th khng phi l bi s ca s nhim sc th n bi, do chng b tha hoc thiu mt hoc vi nhim sc th c th no . l cc th lch bi hay th d bi (aneuploids).
Gim phn I
Gim phn II
Hnh 3.27 S khng phn tch xy ra trong gim phn I (bn tri) v gim phn II (bn phi) vi cc giao t c to ra.
Nguyn nhn ca hin tng lch bi l s khng phn tch (nondisjunction) ca hai nhim sc th tng ng trong qu trnh gim phn hoc nguyn phn. S khng phn tch trong gim phn t n c coi l kt qu ca s kt cp khng ng cch ca cc nhim sc th tng ng trong gim phn sm n ni cc tm ng khng i din nhau trn mt phng k gia, hoc l khng hnh thnh c hnh cho. Kt qu l c hai nhim sc th cng i v mt cc, lm cho mt t bo con tha mt nhim sc th v t bo con kia khng c nhim sc th . Khi cc giao t bt thng (n + 1) v (n 1) ny th tinh vi cc giao t bnh thng (n) s sinh ra cc hp t c b nhim sc th bt thng tng ng: tha mt chic (2n + 1), gi l th ba (trisomy) v thiu mt chic (2n 1), gi l th mt (monosomy). S khng phn tch ph bin nht l trong gim phn I, nhng cng c th xy ra c trong gim phn II (hnh 3.27). Cc
97
kiu t hp nhim sc th khc nh 2n + 2 (th bn: tetrasomy) hoc 2n 2 (th khng: nullisomy) cng c th xy ra, nhng y ta khng quan tm. S khng phn tch cng c th xy ra trong nguyn phn gy ra cc th khm v cc t bo bnh thng v lch bi. Cc th ba c bit n nhiu loi. thc vt, v d in hnh l mt lot cc th ba vi nhng c tnh k l loi c c dc Datura stramonium c Alfred Blakeslee nghin cu vo khong nm 1920. Thc ra, ngi ta pht hin c tt c cc th ba v tng nhim sc th trong s 12 nhim sc th khc nhau loi ny; v mi th ba c mt kiu hnh c trng, th hin r nht l v qu. iu chng t cc nhim sc th khc nhau c cc hiu qu di truyn khc nhau ln tnh trng ny (hnh 3.28a). Cc th ba cng c nghin cu nhiu loi cy trng nh ng, la go v la m nhm xc nh cc nhim sc th mang cc gene khc nhau. hnh 3.28b cn cho thy hiu qu di truyn ca cc th khuyt nhim lin quan 7 nhim sc th khc nhau i vi kiu bng ba ging la m khc nhau (A, B v D).
(a)
(b)
Hnh 3.28 (a) Qu bnh thng ca c c dc Datura (trn cng) v 12 kiu th ba khc nhau, mi kiu c mt v ngoi v tn gi khc nhau. (b) Cc th khuyt nhim lin quan 7 nhim sc th khc nhau ba b gen la m (A, B v D) cho cc hiu qu di truyn khc nhau i vi kiu hnh bng so vi dng bnh thng (hnh cui).
ngi, vic phn tch cc thnh phn nhim sc th ca cc trng hp sy thai t pht cho thy hu nh tt c cc th mt v nhiu th ba u l cc dng gy cht thai. Tuy nhin, mt s trng hp vn c sinh ra vi cc hi chng khc nhau. Ph bin nht l hi chng Down, th ba nhim sc th 21, vi tn s 1/700 s tr s sinh v thng t l vi tui
98
tc ngi m, c bit l t tui 35 tr i (Hnh 3.29). Hi chng Down c m t cch y chng 160 nm, ni chung c cc c trng l tr n v v ngoi chung d thy nh: u to, trn vt, khe mt xch, li hay th ra ..., thng t vong tui 10-40 v him khi sinh sn. C s nhim sc th ca hi chng Down c khm ph u tin vo nm 1959. Kiu nhn ca nhng ngi ny c cho thy cc hnh 3.29c v 3.30a. Danh php hin hnh ch c th c th ba 21 l (47, +21), trong con s 47 ch ton b s nhim sc th v +21 ch ra rng c ba bn sao ca nhim sc th 21. Trong s nhng ngi mc hi chng Down, ch c chng 5% l kiu nhn d hp v chuyn on thun nghch nh ni trc y, v hu ht (khong 95% trng hp) l kt qu ca s khng phn tch trong gim phn. Mt kt qu nghin cu gn y cho thy c im karyotype ca cc trng hp Down nc ta, nh sau: 91% th trisomy 21 thun, 6% th chuyn on nh ni trn v 3% l th khm (Nguyn Vn Rc 2004).
(a)
(b)
(c)
Hnh 3.29 Hi chng Down (a) Mt chu b mc hi chng Down (tri) vi c im cu to bn tay. (b) Mt th v nguy c mc hi chng Down tnh trn 1.000 tr s sinh (trc tung) lin quan vi cc tui ngi m (trc honh). (c) S hnh thnh cc trng tha v thiu NST 21 (tri) v s th tinh gia trng tha NST 21 vi tinh trng bnh thng to ra th ba 21.
99
Cc th ba nhim sc th thng khc l rt him, ch yu l bi v chng khng sng c khi cn l thai nhi. V d, hi chng Patau, th ba 13 (47, +13), c tn s sy thai l 1/33, tn s bt gp l 1/15.000 tr s sinh, v sng cha y su thng. Trng hp khc l hi chng Edward, th ba 18 (47, + 18), tn s bt gp tr s sinh l 1/5.000 v sng khng n mt nm.
(a)
(b)
(c)
Hnh 3.30 Kiu nhn ca mt s th t bin lch bi thng gp. (a) Kiu nhn ngi n mc hi chng Down (47, XX, +21);(b) Kiu nhn ngi mc hi chng Klinefelter (47, XXY). (c) Mt trnh by n gin kiu nhn ca nhng ngi mc hi chng Turner v Kleinfelter.
Ngoi ra, s khng phn tch ca cc nhim sc th gii tnh ngi gy ra mt s kiu nhim sc th bt thng ph bin nh: XO, XXX, XXY v XYY. Chng hn, hi chng Klinefelter XXY (hay 47, XXY) bt gp tr s sinh vi tn s khong 1/2.000; y l trng hp ca nhng ngi nam c cc c im nh v sinh, thn hnh cao khng cn i...
Tai lieu chia se tai: wWw.SinhHoc.edu.vn100

Kiu nhn ca c th mc hi chng Klinefelter c m t hnh 3.30b. Nhng ngi mc hi chng XYY (c th gi l "siu nam"), xut hin vi tn s khong 1/1.000 trong s cc tr nam s sinh; nhng c th ny khng c v g l bnh tt, sc sng v sinh sn bnh thng. Cc nghin cu xa hn cho thy dng nh nhng ngi ny c khuynh hng phm ti, c th c tng quan ti thiu vi hnh vi. Tn s cc c th XYY b ngi t cao ng k so vi cng ng dn c ni chung; tuy nhin, cha ti 5% tng s cc c th XYY l c gio dc dy d ng hong. Hi chng Turner XO (hay 45, X) xut hin tr s sinh vi tn s chng 1/5.000 v tn s sy thai khong 1/18; y l trng hp ca nhng ngi n v sinh, ln khng cn i, c ngn...Cn hi chng "siu n" XXX (hay 47, XXX) bt gp tr s sinh vi tn s khong 1/700; hu nh tt c nhng ngi n mc hi chng ny u v sinh.
Cu hi v Bi tp
1. Th no l chu k t bo? Bng cch no s lng nhim sc th c trng ca mi loi c duy tr n nh trong qu trnh nguyn phn? 2. Bng cch no s lng nhim sc th t trng thi lng bi gim xung cn n bi trong qu trnh gim phn? Ti sao ni cc s kin k trc v k sau ca gim phn I gp phn quan trng trong vic to ra ngun bin d t hp phong ph a dng cc loi sinh sn hu tnh? 3. So snh nguyn phn v gim phn, v cho bit ngha ca chng. 4. Ti sao trong mi t bo s lng nhim sc th (mc n) v hm lng ADN (mc c) l khng ng b trong mi giai on ca phn bo? 5. Hai im sai khc chnh gia s sinh tinh v sinh trng ngi v hu ht cc ng vt c v khc l g? Cc qu trnh pht sinh giao t ng vt v thc vt bc cao c nhng im ging v khc nhau no? 6. Mt sinh vt c s lng nhim sc th lng bi bng 12, c k hiu: Aa, Bb, Cc, Dd, Ee v Ff. (a) C bao nhiu t hp nhim sc th khc nhau c th xut hin trong cc giao t? (b) Xc sut mt giao t nhn c tt c cc nhim sc th vit hoa l g? 7. (a) C th c nhng kiu bin i no xy ra trong cu trc nhim sc th? M t nhng nt chnh xy ra trong mi kiu cng vi hu qu v ng dng ca chng. (b) Gi tn cc trng hp lch bi ph bin ngi v m t cc thnh phn nhim sc th ca chng. 8. Mt rui gim ci thn en mun ee (e = ebony) ng hp c cho

lai vi mt con c hoang di ng hp (e+e+) c chiu tia X. i con c xut hin mt rui ci thn mun. Khi cho rui gim ny lai vi rui gim F1 thu c thn mun: hoang di. Hy gii thch kt qu ny. 9. Gi s rng F1 ca php lai bi tp 8 ta pht hin c hai con rui gim c v ci u c thn en mun. C th ni g v s lng tng i ca i con nu cho hai con rui gim F1 lai vi nhau? (Cho bit cc mt on thng gy cht trng thi ng hp). 10. Cho mt cy d hp ABCDE/abcde lai vi mt cy abcde/abcde, v i con xut hin su dng sau: ABCDE; abcde; Abcde; aBCDE; ABCDe; abcdE. C iu g bt thng trong kt qu ny v bn c th gii thch iu ra sao? (Gi : s thiu vng ca hai hoc nhiu kiu hnh c k vng gi ra mt th d hp o on c lin quan n cc gene lin kt).
Ti liu Tham kho

Ting Vit Dubinin NP. 1981. Di truyn hc i cng (bn dch ca Trn nh Min v Phan C Nhn). NXB Nng Nghip, H Ni. Kimura M. 1983. Thuyt tin ha phn t trung tnh. (Bn dch ca Hong Trng Phn). NXB Thun Ha - Hu, 1993 (tr.17-33). L nh Lng, Phan C Nhn. 1997. C s di truyn hc. NXB Gio Dc, H Ni. Phan C Nhn (ch bin), Nguyn Minh Cng, ng Hu Lanh. 1999. Di truyn hc. NXB Gio Dc, H Ni. Nguyn Vn Rc. 2004. Nghin cu c im karyotype, kiu hnh ca tr Down v karyotype ca b m. Lun n Tin s Y hc, Trng i hc Y H Ni, H Ni. Phm Quang Vinh. 2003. Nghin cu bt thng nhim sc th trong cc th bnh Leucmie cp ngi ln ti Vin Huyt hc Truyn mu. Lun n Tin s Y hc, Trng i hc Y H Ni, H Ni. Ting Anh Campbell NA, Reece JB. 2001. Essential Biology. Benjamin/Cummings, an imprint of Addison Wesley Longman, Inc, San Francisco, CA. Clarke L. 1998. Centromeres: proteins, protein complexes, and repeated domains at centromere of simple eukaryotes. In: Current Opinion in Genetics & Developmment, Vol 8, No2 (Allis CD and Gasser SM, eds.), pp 212-218.

Clegg CJ, Mackean DG. 2000. Advanced Biology: Principles and Applications. 2nd ed, John Murray Published Ltd, London. Dobie KW, Hari KL, Maggert KA, Karpen GH. 1999. Centromere proteins and chromosome inheritance: a complex affair. In: Current Opinion in Genetics & Developmment, Vol 9, No2 (Kadonaga JT and Grunstein M, eds.), pp 206-217. Hartl DL, Freifelder D, Snyder LA. 1988. Basic Genetics. Jones and Bartlett Publishers, Inc, Boston - Portola Valley. Lewis R. 2003. Human Genetics: Concepts and Applications. 5th ed, McGraw-Hill, Inc, NY. Li W-H. 1983. Evolution of duplication genes and pseudogenes. In: Evolution of Genes and Proteins (Nei M, Koehn RK, eds.), pp. 14-37. Sinauer Associates Inc.-Publishers, Sunderland, Massachusetts, USA. Biggins S. and Murray AW. 1999. Sister chromatid cohesion in mitosis. In: Current Opinion in Genetics & Developmment, Vol 9, No2 (Kadonaga JT and Grunstein M, eds.), pp 230-236. Russell PJ. 2003. Essential Genetics. Benjamin/Cummings Publishing Company, Inc, Menlo Park, CA. Schrock E, du Manoir S, Veldman T, Schoell B, Weinberg J, FergusonSmith MA, Ning Y, Ledbetter DH, Bar-Am I, Soenksen D, Garini Y, Ried T. 1996. Multicolor spectral karyotyping of human chromosomes. Science 273: 495. Verma RS and Babu A. 1995. Human chromosomes: Principles and Techniques. 2nd ed., McGraw-Hill, Inc, NY. (Ch2-6, pp.6-231). Weaver RF, Hedrick PW. 1997. Genetics. 3rd ed, McGraw-Hill Companies, Inc. Wm.C.Browm Publishers, Dubuque, IA. Mt s trang web http://www.mhle.com/lewisgenetics5 http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=OMIM http://www.genetic.org http://www.fivepminus.org http://www.ndss.org http://www.members.aol.com/cdousa/cdo.htm http://www.turner-syndrom-us.org http://www.trisomy.org

Chng 4
Di truyn hc Nhim sc th
Nh tho lun cc chng trc, nguyn l phn ly c lp ca Mendel ch nghim ng trong trng hp cc gene nm trn cc nhim sc th khc nhau v c l gii bng s phn ly ngu nhin ca cc nhim sc th trong gim phn. Tuy nhin, vo u thp nin 1910, Thomas Hunt Morgan (hnh 4.1a) da vo cc kt qu nghin cu rui gim Drosophila melanogaster (hnh 4.1b) nhn ra rng c nhiu gene cng nm trn mt nhim sc th. iu khng nh ng n ny sm b sung v lm sng t cho cc nguyn l di truyn Mendel, ng thi t nn tng vng chc cho s pht trin ca di truyn hc trong sut na u th k XX. Trong chng ny, chng ta ln lt tm hiu thuyt di truyn nhim sc th vi cc vn xc nh gii tnh, cc kiu di truyn lin kt i vi hai hoc nhiu gene trn cng mt nhim sc th, cng nh cc phng php kinh in dng lp bn di truyn cc sinh vt.
I. Trng phi Morgan vi thuyt di truyn nhim sc th

T 1910, Morgan cng vi ba cng s l Alfred H.Sturtevant, Calvin Bridges v Herman J. Muller (hnh 4.1c) xy dng thnh cng thuyt di truyn nhim sc th (chromosome theory of inheritance) da trn i tng nghin cu ni ting l rui gim D. melanogaster. Trc tin, chng ta hy tm hiu c im ca i tng, phung php nghin cu v ni dung ca hc thuyt ny.
(a)
(b)
(c)
Hnh 4.1 (a) T.H.Morgan; (b) Rui gim D. melanogaster, i tng nghin cu ni ting ca Morgan; v (c) Herman Muller, mt trong ba mn xut sc ca Morgan vi phng php gy t bin bng tia X.
1. Tm quan trng ca rui gim Drosophila 1.1. Cc c im v gi tr ca rui gim trong nghin cu di truyn hc Drosophila melanogaster (hnh 4.1 v 4.2) c l l sinh vt ni ting

nht c dng lm sinh vt m hnh (model organism) cho cc nh di truyn hc. Rui gim thuc lp cn trng (Insecta), b hai cnh (Diptera). Chng rt thch mi ln men ca cc h da vi c v c bit l nhng tri cy chn mui nh chui, mt hay cam, chanh..., v vy chng c bit di ci tn thng dng l rui gim hay "rui tri cy" (fruit-flies). Rui gim phn b rng khp cc vng n i v nhit i trn hnh tinh chng ta.
(a)
(b)
Hnh 4.2 (a) S khc nhau v hnh thi ngoi gia rui gim c (trn) v rui gim ci (di); v (b) b nhim sc th lng bi 2n = 8 ca chng, vi cp nhim sc th gii tnh XY- con c (tri) v XX- con ci.
Gi tr ca rui gim Drosophila trong cc th nghim di truyn nm trong cc c im sau (cc hnh 4.2, 4.3 v 4.4): - Mi cp rui gim sinh c hng trm con trong mt la; - Vng i ngn, ch c hai tun l l chng c th nhanh chng t ti tui trng thnh tham gia sinh sn; v chu k sng c th rt xung cn 10 ngy, nu nhit 25oC. Cc rui ci trng thnh v mt sinh dc ni trong 12 gi, v chng li trng ha nhng trong hai ngy.. - Sau khi giao phi, cc con ci c th bo qun cc tinh trng, v vy cn thit phi tin hnh cc php lai vi cc con ci ang cn trinh (virgin). Rui ci cn trinh c th d dng nhn ra qua mu ct su (meconium) hay xm nht ca c th chng (hnh 4.3d) v phn nhng di dng chm en c th nhn thy xuyn qua vng bng. - Rui gim Drosophila tng i d nui, v d dng phn bit cci cc giai on non v trng thnh cch ly v tin hnh lai. Bn cnh s phn bit ngoi hnh cc rui non cn trinh nh ni trn, giai an trng thnh rui c thng khc vi rui ci cc im sau: c th b hn; vng bng di c ba vch en vi vch di cng rng, trong khi rui ci c nm vch ri nhau; chm bng con c hi trn v con ci nhn (hnh 4.2a). - B nhim sc th y ca cc t bo soma rui gim ch c bn cp, 2n = 8 (hnh 4.2b). Ton b b gene ca Drosophila c xc nh trnh t trong thi gian gn y.
(a)
(b)
(c)
(d)
(e)
(f)
(g)
(h)
(i)
(j)
(k)
(l)
Hnh 4.3 Mt s th t bin quan trng ca rui gim Drosophila. Ch thch: (a-b) th t bin mt trng v mt kiu di; (c) con c trng thnh (tri) v con c cn trinh c mu ct su; (d) con ci trinh c mu ct su; (e) cc dng t bin khc nhau v cnh; (f) th t bin ny l mt v d v hai kiu hnh cnh ngn/thn mu en mun; (g) th t bin cnh qun con ci (tri) so vi con ci bnh thng; (h) th t bin hai t ngc; (i) th t bin antennapedia- kiu chn ru v (j) anten kiu di; (k) so snh th t bin anten (di) v dng bnh thng (trn); (l) Mt kiu di vi cc dng mt thi (Bar eye) d hp v ng hp (theo th t t trn xung).
- Rui gim c nhiu tnh trng, c bit l cc th t bin t pht nh mt trng hoc c to ra trong phng th nghim (nh thn en, cnh ngn, cnh qun, mt nu...); cc tnh trng ny c th phn bit bng mt thng, knh lp hoc knh hin vi quang hc (hnh 4.3 v 4.5). - Cc t bo tuyn nc bt ca u trng rui gim (hnh 4.4) c cha cc nhim sc th khng l a si (nh gii thiu chng 3); y l im thun li cho vic xc nh cc phn c th ca cc nhim sc th. Cc bng ny t chng khng phi l cc gene nhng rt hu ch cho vic lp bn cc gene trn cc nhim sc th.

Hnh 4.4 Cc a mm (trong u trng) t hnh thnh nn cc c quan ca rui gim trng thnh. T trn xung: cc phn ph ming, mnh trn v mi trn, anten, mt, chn, cnh v c quan sinh dc.
Ngoi ra, cc dng c v ha cht c s dng trong cc th nghim rui gim l tng i n gin, c gii thiu hnh 4.5 di y.
(a)
(b)
(c)
Hnh 4.5 Cc dng c th nghim vi rui gim. (a) ng nghim nui v lai rui gim; (b) khay ng cc l ha cht v mt s vt dng khc; v (c) knh hin vi quang hc dng cho nghin cu hnh thi v t bo hc.
1.2. Cc tnh trng c kim sot v mt di truyn ca rui gim Mt qun th bnh thng ca rui gim Drosophila bao gm cc con rui in hnh c thn xm, ccnh di v mt . Dng rui gim ny l ph bin nht, v c cc nh di truyn hc xp vo kiu di (wild type). Nhiu dng t bin cng c pht hin trong t nhin. Mt con rui t bin c mt xut pht im di truyn t nht l mt trong s cc tnh trng ca dng rui gim bnh thng. Cc tnh trng t bin c bit n bng mt tn gi. V d: t bin tn gi k hiu c tnh mt trng trng (white) w ln thn en mun mun (ebony) e ln cnh ngn ngn (vestigial) vg ln mt nu nu (brown) bw ln mt thi (Bar) Bar B tri Cc th t bin ln c k hiu bng cc ch ci vit thng, v cc tnh trng tri tng ng c biu th bng cc ch ci vit hoa.

Trong cc th nghim lai di truyn, allele bnh thng mt locus c th c cc nh di truyn hc trc y k hiu bng du "+". K hiu ny thng c b qua mc nhp mn, mc d n thng c dng cho rui gim. Tuy nhin, i vi bc i hc, cc alelle bnh thng c quy cho "kiu di" v s dng cc k hiu ca cc nh di truyn hc. 2. Thuyt di truyn nhim sc th Rui gim Drosophila melanogaster c nh di truyn hc ngi M (USA), T.H. Morgan (1866-1945), s dng trong nghin cu di truyn hc t nhng nm u ca th k XX, trong khi ang lm vic ti Hc vin Cng ngh California (California Institute of Technology). Nh s dng rui gim ny, Morgan v cc cng s ca mnh xy dng thnh cng hc thuyt di truyn nhim sc th (chromosome theory of heredity). Trc tin, ta hy tm lc cc ni dung chnh ca thuyt di truyn nhim sc th v nhng ng gp ca trng phi Morgan cho s pht trin ca di truyn hc. (1) Hc thuyt ny xc nhn rng: gene l n v c s ca tnh di truyn nm trn nhim sc th. Trn mi nhim sc th c nhiu gene phn b thng hng, mi gene chim mt v tr xc nh gi l locus; cc gene trn mi nhim sc th hp thnh mt nhm lin kt. S nhm lin kt gene chnh l s nhim sc th n bi, cn gi l b gene (genome). (2) Trong qu trnh gim phn, cc gene trn cng nhim sc th c xu hng phn ly cng nhau v giao t. y l c s ca hin tng di truyn lin kt gene hon ton v di truyn lin kt (linkage) ni chung. (3) Tuy nhin, trong qu trnh gim phn c th xy ra s trao i cho (crossing over) mt s on gia hai nhim sc th tng ng, ko theo s trao i cc gen gia chng, cn gi l ti t hp hay hon v gene. y chnh l c s ca hin tng lin kt gene khng hon ton. (4) Tn s ti t hp (rate of recombination) ca cc gene l mt s hu t, tha mn gii hn t 0,0 n 0,5 (tc khng vt qu 50%). i lng ny t l thun vi khong cch gia cc gene v ph thuc vo mt s yu t khc nh gii tnh cng nh mc c ch bi cc trao i cho ng thi ti nhiu im trn mt cp tng ng. Da vo cc tn s ti t hp gene ny ta c th thit lp bn nhim sc th (chromosome map) ca cc loi, hay cn gi l bn lin kt (linkage map). (5) Vn cc nhim sc th xc nh gii tnh (sex-determining chromosomes) v hin tng di truyn lin kt vi gii tnh (sex linkage) c lm sng t ln u tin bi hc thuyt ny. (6) Ngoi ra, trng phi ca Morgan xc nh rng: gene - n v

di truyn hc then cht ny ng ba vai tr: (i) Gene l n v chc nng, ngha l gene c xem nh mt th thng nht ton vn kim sot mt tnh trng c th. (ii) Gene l n v ti t hp, ngha l gene khng b chia nh bi s trao i cho (v theo quan im ny, trao i cho khng xy ra bn trong phm vi mt gene m ch xy ra gia cc gene); nh th gene c coi l n v cu trc c s ca vt cht di truyn, nhim sc th. (iii) Gene l n v t bin, ngha l nu t bin xy ra trong gene d bt k v tr no hoc vi phm vi ra sao, ch gy ra mt trng thi cu trc mi tng ng vi mt kiu hnh mi, kiu hnh t bin, khc vi kiu hnh bnh thng. Tuy nhin, quan nim ny vn cn cha r rng v khng thc s chnh xc theo quan im ca di truyn hc hin i (s c tho lun chng 6). Trong vic xy dng hc thuyt di truyn nhim sc th, phi k n nhng ng gp to ln ca cc mn xut sc ca Morgan, l: Alfred H.Sturtevant vi vic xut phng php xy dng bn di truyn nm 1913; Calvin Bridges vi vic pht hin ra c ch xc nh cc kiu hnh gii tnh rui gim nm 1916; v Herman J.Muller vi s pht trin phng php gy t bin bng tia X nm 1927. Nh ng gp to ln , Morgan c trao gii thng Nobel nm 1933 v Muller - nm 1946.
II. S xc nh gii tnh (sex determination)

C ch t nhin m trong mt c th ca mt loi phn tnh (dioecious species) tr thnh con c hoc con ci (hay lng tnh, hermaphroditic) c gi l xc nh gii tnh (sex determination). Thc ra, khng c mt phng thc ph bin no cho vic xc nh gii tnh; nhng cho n nay, cc nh sinh hc khm ph ra nhiu c ch xc nh gii tnh, tp trung vo hai tiu ch sau (xem cc Bng 4.1- 4.3). mt s loi, gii tnh c xc nh sau khi th tinh bi cc nhn t mi trng nh nhit hoc s c mt ca cc th km gii tnh. Kiu xc nh gii tnh ny c gi l xc nh gii tnh do mi trng (environmental sex determination = ESD). cc loi khc, gii tnh c xc nh lc th tinh bng s t hp ca cc gene m hp t nhn c. Kiu xc nh gii tnh ny c gi l xc nh gii tnh do kiu gene (genotypic sex determination = GSD). Di y chng ta ln lt xt qua hai h thng ny (v chi tit, xem trong: Kalthoff 1997, tr.660-685; v Yablokov 1986, tr.17-30). Ring vn bt hot nhim sc th X s c tho lun mc II-3-2, v n c lin quan n di truyn lin kt gii tnh. 1. S xc nh gii tnh do kiu gen (GSD) hu ht cc sinh vt, gii tnh c xc dnh bng s sai khc nhim sc th v cc gene trn chng (bng 4.1). V d u tin v s xc nh

gii tnh do nhim sc th c ghi nhn rp Protenor nm 1905, trong cc con ci c khm ph c hai nhim sc th X v con c ch c mt X. Cc giao t c c th c mt X hoc khng c nhim sc th X c bit l c t l ngang nhau, v cc giao t ci th lc no cng mang mt X, v vy s lng i con c XX (ci) v X (c) l ngang nhau. Sau khng lu, nhiu nh nghin cu xc nh s c mt ca mt nhim sc th gii tnh Y cc sinh vt khc. cc loi ny, s c mt ca Y cng vi mt X cho ra con c, trong khi hai bn sao ca X sinh ra mt con ci. Vi li, cc giao t c gm hai kiu, v vy c s lng tinh trng hay ht phn mang X v mang Y bng nhau. Trong trng hp gii tnh c c hai kiu nhim sc th gii tnh nh th, th kiu XY c gi l gii tnh d giao t (heterogametic sex), cn kiu XX c gi l gii tnh ng giao t (homogametic sex). hnh 4.6 cho thy cc nhim sc th X v Y, v c ch xc nh gii tnh ngi. mt s sinh vt thuc cc lp nh chim v b st th ngc li, cc con c (trng) l ng giao t v cc con ci (mi) l d giao t. trnh s nhm ln, ngi ta thng gi cc nhim sc th gii tnh ny l Z v W; nh vy cp nhim sc th gii tnh con c l ZZ v con ci l ZW. Kt qu l, cc con ci cho hai loi giao t (mt loi mang Z v mt loi mang W vi t l ngang nhau), trong khi cc con c ch cho mt loi giao t tt c u mang mt nhm sc th Z. S hiu bit v gii tnh ny cng vi s di truyn lin kt gii tnh c ngha thc tin to ln trong chn nui gia cm. Vn ny s c tho lun tr li cui mc III.
Bng 4.1 Cc kiu xc nh gii tnh khc nhau bi nhim sc th Sinh vt Hu ht ng vt c v, mt s cn trng (tt c bn hai cnh), mt s c, mt s thc vt Rp Protenor, mt s cn trng khc, kangoroo Lp chim, hu ht cc b st, bm m B cnh mng (Hymenoptera) Con ci XX XX ZW Lng bi Con c XY X ZZ n bi
Hu nh tt c cc ng vt c v, s c mt ca mt nhim sc th Y cn thit cho s pht trin ca kiu hnh ging c. Chng hn, nhng ngi mc hi chng Turner (45, X) u l n. Hn na, nhng ngi mc hi chng Klinefelter (47, XXY hoc 48, XXXY) u l nam mc d h c th c ti hai hoc ba nhim sc th X. iu chng t nhim sc th Y c cha gene xc nh tnh c (maleness). Ngy nay ta bit r rng chnh l nhn t xc nh tinh hon (TDF = testis-determining factor) nm trn vai ngn nhim sc th Y (hnh 4.6c). Sinclair v cs (1990) gi

n l gene SRY (sex-determining region Y).
vng gi NST thng
vai p vai q vng gi nhim
(a)
(b)
(c) sc th thng
Hnh 4.6 (a) Cc nhim sc th X v Y ng vt c v, i din l ngi. (b) C ch xc nh gii tnh ngi. (c) Nhim sc th Y ca ngi vi gene SRY nm k vng gi nhim sc th thng u mt ca vai ngn.
Cn lu rng, gn y ngi ta cng xc nh c gene "chuyn i" (switch gene) SRY trn nhim sc th Y bng cch kim tra mt vi c th him hoi b o ngc gii tnh (sex-reversed), ngha l con c XX v con ci XY. Cc con c XX thc t c mang mt mu ca Y cha gene xc nh tnh c, cn cc con ci XY th li thiu hn vng nh vy Y. Chng hn, mt vi bnh nhn o ngc gii tnh vn l nhng ngi nam v sinh r rng l mang c hai nhim sc th X v khng c Y, v mt s ngi n mc hi chng Turner th li mang mt X v mt Y r rng. Cc kt qu phn tch k hn (trn bn ngi bnh o ngc gii tnh nh p dng cc vt d DNA c to dng t nhim sc th Y vo Southern blots ton b DNA b gene ca h) cho thy rng hu ht cc trng hp ny u c lin quan n s chuyn mt on DNA c th trn nhim sc th Y (nm st vng gi nhim sc th thng pseudoautosomal region); v chnh s chuyn on ny gy ra hin tng o ngc gii tnh. iu ny c gii thch l do cc s kin trao i cho him hoi gia X v Y trong gim phn ngi nam hay con c. Lu rng gene SRY+ m ha mt protein bm DNA v n c bo tn cao cc ng vt c v. John Gubbay v cs (1990) pht hin ra mt gene tng ng chut, k hiu l Sry+, khng c mt trong chut ci XY b o ngc gii tnh. rui gim, c mt th t bin autosome ca gene "chuyn i", tra (transformation), m khi trng thi ng hp trong cc c th XX bin i chng thnh nhng con c, ch khng phi l cc con ci nh k vng. Tnh hung ngc li c bit ngi, trong cc c th mang mt t bin trn nhim sc th X c mt trng thi gi l n ha tinh hon (testicular feminization). Trong trng hp ny, cc c th XY v mt kiu hnh l n, c ngc n nang v m o, nhng

h u v sinh. t bin ny hin nhin l c lin quan ti mt cht mang hormone m n ngn khng cho nhng ngi ny tit ra testosterone; v hu qu l h khng pht trin c cc tnh trng nam th cp.
Bng 4.2 T l X/A v cc kiu hnh gii tnh ca rui gim D. melanogaster (theo Bridges 1921, trch t Yablokov 1986) B nhim sc th (*) 3X : 2A 4X : 3A 4X : 4A 3X : 3A 2X : 2A 1X : 1A 3X : 4A 2X : 3A 1X : 2A 2X : 4A 1X : 3A T l X/A 1,50 1,33 1,00 1,00 1,00 1,00 0,75 0,67 0,50 0,50 0,33 Kiu hnh gii tnh Siu ci Siu ci Ci t bi Ci tam bi Ci lng bi Ci n bi Gii tnh trung gian Gii tnh trung gian c lng bi c t bi Siu c Ghi ch Bt th, tnh trng thin v ci Bt th, tnh trng thin v ci Hu th hu th gim Hu th Bt th Bt th Bt th Hu th Hu th Bt th, tnh trng thin v c
(*) X - s nhim sc th X, A - s b n bi nhim sc th thng.
Mc d rui gim Drosophila nhng con ci l XX v con c l XY, song s c mt ca Y khng nht thit cho kiu hnh c (nh ng vt c v). Chng hn, cc kiu hnh ngoi l ny c Calvin Bridges, mt hc tr xut sc ca Morgan, pht hin t nm 1916 l cc con c XO c mt v cc con ci XXY mt trng. Sau , Bridges pht trin cc ni rui gim tam bi. Khi kim tra cc c th c s lng nhim sc th X khc nhau, ng pht hin ra rng kiu hnh gii tnh rui gim l mt hm s ca t l gia s lng cc nhim sc th X (X) v s lng cc b nhim sc th thng (A). V d, bng 4.2 cho thy cc con ci bnh thng c hai nhim sc th X v hai b nhim sac th thng, ngha l t l X/A = 2/2 = 1. Cc con c bnh thng c mt X v hai b nhim sc th thng, do X/A = 1/2 = 0,5. Cng vy, cc con c v ci ngoi l vn c cc t l tng ng l 0,5 v 1,0. Khi Bridges kim tra cc con rui c ba nhim sc th X v hai b nhim sc th thng, tc 3/2 = 1,5, ng thy rng chng u l cc con ci bt dc (i khi gi l "siu ci", metafemales), trong khi cc con rui c mt X v ba b nhim sc th thng, X/A = 1/3 = 0,33, u l cc con c bt dc hay gi l dng siu c (metamales), Cui cng, cc con rui c hai X (XX) v ba b nhim sc th thng, X/A = 2/3 = 0,67 v chng c cc c im kiu hnh ca c hai gii v c gi l gii tnh trung gian (intersexes). T cc pht hin ny, Bridges ngh rng, nhn chung, cc nhim sc th X

xc nh gii ci v cc nhim sc th thng (autosomes) xc nh gii c. Ni cch khc, khi t l X/A l 1 hoc ln hn, cc rui gim s c ku hnh ci, v khi t l X/A l 0,5 hoc nh hn th chng s c kiu hnh c. Tuy nhin, ta cn lu rng mc d thm ch nhim sc th Y khng nht thit cho kiu hnh c rui gim, nhng n nht thit cn cho hu th (cc con c XO v cc con rui siu c u bt dc). Ngoi ra, cc sinh vt thuc b cnh mng (Hymenoptera) nh ong mt v ong bp cy, chng khng c nhim sc th gii tnh; trong trng hp ny, nh ni chng 3, con ci l lng bi v con c l n bi. V d, ong mt (Apis mellifera), cc ong ci bao gm c ong cha ln ong th l lng bi (2n = 32) v cc ong c - n bi (n = 16). Khi gim phn, ong cha cho cc giao t ci (n = 16), v ong c vi hin tng lng bi gi gim phn cho cc giao t c (n = 16). Cc trng c th tinh (2n = 32) pht trin thnh cc ong ci, ch mt vi con c u tin mm sa cha lin tc trong 3-5 ngy u s pht trin thnh ong cha; cn cc trng khng c th tinh s pht trin thnh cc ong c. 2. S xc nh gii tnh do mi trng (ESD) Mt trng hp ni bt nht v xc nh gii tnh do mi trng l Bonellia viridis, mt loi u trng giun bin (Leutert 1975). Cc con ci trng thnh ca loi ny bm vo cc mm i dng. C th con ci di hn 10 cm, con c rt b (1-3 mm) sng k sinh bn trong con ci. Cc u trng ca Bonellia sng nh l thnh phn ca sinh vt tri ni (plankton), ngha l, cc sinh vt nh b di chuyn th ng trong nc. S xc nh gii tnh xy ra khi cc u trng c th tinh trn mt gi th v bin thi. Nhng u trng c th tinh trong s cch ly (sng t do) th pht trin thnh cc con ci, trong khi cc u trng chui vo trong mt con ci trng thnh th tr thnh cc con c. Mt s trng hp khc, chng hn nh cc trng c su chu M (Alligator) c a vo nhit cao cho ra hu ht l con c, cn nu nhit thp cho ra hu ht l con ci. ra th ngc li (xem bng 4.3).
Bng 4.3 Cc kiu xc nh gii tnh khc nhau do mi trng (ESD) Loi Ra (Turtles) C su (Alligators) Meloidogyne incognita Bonellia viridis C ch Nhit Nhit Mt qun th S c mt con ci Cc gii tnh m: ci Lnh: ci Phn tn: ci C: c Lnh: c m: c Tp trung: c Khng: ci
Ngun: Hodgkin (1992), trch chn theo sa i ca Kalthoff (1997, tr.662).

Ngoi ra, gii tnh ca mt s loi c chu nh hng bi u th sng thnh n, cn cc loi thc vt no li cho cc gii tnh khc nhau ty thuc vo di ngy hoc cc nhn t khc nh hng ln tc sinh trng ca chng. Tm li, cc loi sinh sn hu tnh giao phi, ni chung t l c/ci trn quy m qun th-loi l xp x 1:1, v c th gii thch bng c ch phn ly ca cc nhim sc th gii tnh v cc giao t trong qu trnh gim phn v s kt hp ngu nhin ca chng trong th tinh.
III. S di truyn lin kt vi gii tnh (sex-linked inheritance)

Trong cc chng 1 v 2 chng ta mi ch cp n cc gene trn nhim sc th thng (autosomes), tc cc nhim sc th tn ti trong cc t bo soma ng ngha l cc cp tng ng; v vy m cc kt qu lai thun nghch l tng ng nhau. Tuy nhin, mt s th nghim ca Morgan rui gim vo nm 1910 cho thy cc kt qu lai thun nghch (reciprocal crosses) l khc nhau. Di y ta xt mt s trng hp di truyn lin kt vi gii tnh ch yu rui gim v ngi, mi lin quan gia di truyn lin kt gii tnh v s bt hot ca nhim sc th X, cng nh s di truyn ca mt s tnh trng b gii hn bi gii tnh v chu nh hng ca gii tnh. 1. c im di truyn ca cc gen trn nhim sc th X v Y Bn cht ca cc nhim sc th gii tnh X v Y l khc nhau (hnh 4.6). Trong khi nhim sc th X thng rt ln v mang nhiu gene kim sot ch yu cc tnh trng thng, th nhim sc th Y li rt b v cha t gene. V vy s di truyn lin kt vi gii tnh (sex-linked inheritance), tc s di truyn ca cc gen trn cc nhim sc th gii tnh cng khc nhau. 1.1. S di truyn lin kt - X Hin tng di truyn lin kt gii tnh c Morgan khm ph ln u tin rui gim Drosophila. Bnh thng mu mt ca rui gim c mu . Trong mt qun th Drosophila nui qua nhiu th h, Morgan pht hin mt th t bin mt trng rui c (xem hnh 4.3a-b). Khi cho lai gia rui c mt trng ny vi rui ci mt thun chng, i con F1 nhn c tt c mt (ng nh k vng allele mt trng l ln). Sau khi cho tp giao cc rui F1 vi nhau, F2 thu c 2.459 rui ci mt , 1.011 rui c mt v 782 rui c mt trng. Nhn chung, t l : trng khng gn vi t l 3:1. Tuy nhin, kt qu bt ng l tt c cc con rui mt trng u l c (hnh 4.7a)! Hn na, khi cho rui c mt trng lai vi rui ci mt F1, i con sinh ra gm c hai loi mt v trng mi gii tnh vi t l tng ng. iu chng t tnh trng mt

trng phn b c hai gii, ch khng phi ch gii c. Mt khc, khi tin hnh php lai nghch gia rui ci mt trng F2 trong th nghim ni trn vi rui c mt , kt qu thu c khc vi php lai thun trc y, tt c rui ci mt v tt c rui c mt trng (hnh 4.7b).
Tinh trng
Trng
Ci d hp mt
c bn hp t mt trng
(a) (b) Hnh 4.7 Cc php lai thun nghch v mt trng v mt rui gim. (a) Php lai thun: P = ci mt c mt trng F1 tt c F2 3 : 1 trng (mt trng ch c gii c); v (b) Php lai nghch: P = ci mt trng c mt F1 gm tt c rui ci mt v tt c rui c mt trng.
Vi cc kt qu , Morgan cho rng cc tnh trng mt v mt trng ny di truyn lin kt gii tnh, do cc allele tri (w+) v (w) ln tng ng nm trn nhim sc th X kim sot; con ci l XX v con c l XY. V thut ng bn hp t (hemi-zygote) c ng dng m t trng hp ny. Ta c th tm tt cc c im ca s di truyn lin kt X nh sau: (1) Mt c th ci d hp t s truyn allele ln cho mt na con ci v mt na con c ca n. (2) Allele ln con c tn ti trng thi bn hp t s biu hin ra kiu hnh ln (nn kiu hnh ln ny ph bin gii c), v con c s truyn allele ln ny cho cc con ci (female) ca n. (3) Hin tng di truyn allele ln t b cho con gi v biu hin chu ngoi

trai nh th r rng l c s cch qung th h, v n c gi l di truyn cho. (4) Ni chung, vic nhn bit mt tnh trng no tun theo quy lut di truyn lin kt-X c th da vo cc t l kiu hnh khc nhau t cc php lai thun nghch, hoc s phn b khng u ca cc kiu hnh hai gii, hoc bng cch s dng "php th c lp" (quy tc nhn) nh ni chng 1. By gi ta xt s qua s di truyn lin kt gii tnh (cc gene trn X) ngi. Nm 1994, McKusick lit k 161 locus c xc nh trn nhim sc th X ngi. Tuy nhin, theo thng k ca OMIM, tnh n ngy 8/2/2005 con s ny l 539 (OMIM 2005) trong tng s hn 1.400 gene cha trong 150 triu cp base ca nhim sc th ny (NCBI 2005). D nhim sc th X mang hng trm gene nh vy nhng rt t gene ny, nu c, trc tip tc ng ln gii tnh. Tuy nhin, nh ni, s di truyn ca cc gene ny tun theo cc quy tc c bit, v: (i) ngi nam ch c mt nhim sc th X n c; (ii) hu nh tt c cc gene trn X khng c cc gene tng ng trn Y; do vy (iii) bt k gene no trn X s c biu hin nam gii, d n l ln n gii. Nhng ngi n d hp t c gi l "th mang" (carrier) v mc d h khng cho thy cc triu chng, nhng li truyn gene ny cho khong mt na s con trai (s pht trin bnh) v mt na cho con gi ca h (cng l cc th mang). Mt v d quen thuc v s di truyn lin kt-X l bnh mu kh ng (hemophilia) do mt allele ln ca gene hemophilia A nm gn u mt vai di gy ra (hnh 4.8). Cc c th b bnh ny mt kh nng tng hp mt protein (nhn t VIII) cn thit cho s ng mu bnh thng. Mt ph h ni ting ca bnh ny bt u t gia nh n hang Victoria nc Anh (England). Mt trong s cc con trai ca n hang b bnh mu kh ng, v hai trong s cc c con gi ca b l cc th d hp t v gene ny ( sinh ra
Gene SRY (TDF)
Hnh 4.8 Bn nhim sc th X ca ngi (bn tri) cho thy v tr mt s gene bnh, v s khng tng ng gia n vi nhim sc th Y.

ba chu ngoi trai ca b u mc bnh mu kh ng v bn chu ngoi gi tt c u d hp t). Cc allele ln t hai trong s bn c gi (chu ngoi) d hp t ny c nhp vo cc gia nh hong tc ca Nga v Ty Ban Nha. Cn gia nh hong tc nc Anh hin gi do pht xut t mt hong t bnh thng ca n hong nn khng mc bnh ny. Hnh 4.9 cho thy gia nh ca Czar Nicholas II nc Nga (Russia), trong ngi v Alexandra chnh l chu ngoi gi ca n hong Victoria. Cu con trai Alexis ca h (pha trc hnh) mc bnh hemophilia v b hnh hnh tui 14. Trong khi bn c con gi ca h u bnh thng. i vi bnh hemophilia, cc kiu gene v kiu hnh ca nhng ngi n v nam thng c biu din nh sau:
N
Kiu gene Kiu hnh
Nam
Kiu gene
Kiu hnh
X X XHXh XhXh
bnh thng th mang bnh
X Y XhY
bnh thng bnh
Bnh hemophilia ch nh hng n 0,004% (hay 1/25.000) trong s nam gii ca qun th, ngha l tn s allele ny bng 0,00004. Vy xc sut cho mt ngi n mc bnh ny l khong (0,00004)2.
Hnh 4.9 Mt bc nh ca gia nh Hong Czar Nicholas II. V ng, Czarina Alexandra, ngi bn tri, vi bn c con gi tt c u c mu ng bnh thng, v ring mi cu con trai Alexis (pha trc) b bnh hemophilia.
Mt trng hp khc kh ph bin l bnh m mu -lc (red-green colour blindness). Nhng ngi mc bnh ny khng th phn bit cc mu m nhng ngi khc nhn thy nh mu lc, vng, cam v (chng c nhn thy nh l mt mu). pht hin dng bnh ny ngi ta s dng cc s mu chng hn nh Phiu trc nghim m mu ca Ishihara (Ishihara's tests for colour blindness). Bnh ny do mt allele ln trn X (nm gia hai gene hemophilia A v B) gy ra; nam gii c khong 8% mc bnh ny trong khi n ch khong 0,4%. Ngoi ra, ri lon dng c Duchenne (Duchenne muscular distrophy = DMD) l mt cn bnh qui c thm thng m hu ht xy ra tr em, cuc i chng gn cht vi chic xe ln, ht th cng kh khn. Hin gi

mt ngi bnh ny cng him khi sng st qu 20 tui. Bnh DMD nh hng ti 1 trn 4.000 tr em nam, v khong 1/3 trong s l hu qu ca cc t bin t pht trong gene ny. Gene DMD nm trn vai ngn ca nhim sc th X (xem hnh 4.8). Theo s liu hin gi, gene DMD l gene c kch thc ln nht, khong 2,4 triu cp base, tng ng khong 1,5% chiu di ca nhim sc th X (2,4x106 : 150 106 = 0,016). Trc khi kt thc phn ny, ta hy cp mt vi ng dng thc tin ca s hiu bit v di truyn gii tnh v lin kt vi gii tnh. Trong chn nui gia cm, vic xc nh gii tnh lc chng cn non hoc thm ch giai on trng tht l quan trng. Chng hn, g, mt gene trn nhim sc th Z c mt allele ln xc nh mu lng vng (ZsZs hoc ZsW), v mt allele tri xc nh lng bc (ZSZS, ZSZs hoc ZSW). Khi cho lai gia con mi lng bc (ZSW) v con trng lng vng (ZsZs) s cho i con tt c con trng lng bc (ZSZs) v tt c con mi lng vng (ZsW). R rng l, vi hiu bit quy lut di truyn cho, cho php ta d dng xc nh gii tnh khi g con mi mt ngy tui. Bn hy kim tra php lai ngc li (chng hn ZSZS ZsW) xem th liu php lai ny c th dng xc nh gii tnh ca cc g con mi n hay khng? 1.2. S di truyn lin kt-Y Ni chung, nhim sc th Y rt b, cha t gene. Nhim sc th Y rui gim hu nh khng mang gene, trong khi ngi c 48 gene bit trnh t trong tng s hn 200 gene c c tnh l cha trong khong 50 triu cp base (OMIM-NCBI 2005). hai u mt ca nhim sc th Y c hai vng c gi l cc vng gi nhim sc th thng (pseudoautosomal regions; hnh 4.6c) bi v cc gene nh khu bn trong chng (cho n nay ch pht hin c 9 gene) u c di truyn ging nh bt k cc gene no thuc nhim sc th thng. V s trao i cho gia X v Y ch c th xy ra hai vng tng ng rt nh ny ca Y. Mc d 95% ca nhim sc th Y nm gia cc vng gi tng ng, nhng s gene c pht hin y l cha ti 80. Mt s gene ny m ha cc protein dng chung cho tt c cc t bo (v c hai gii tnh). Nhng gen cn li m ha cc protein hnh nh ch hot ng trong cc tinh hon. Gene ch cht nht nhm sau l gene xc nh tinh hon TDF, hay gene SRY, nh khu trn vai ngn ngay bn ngoi vng gi nhim sc th thng (hnh 4.6c). Cc tnh trng c quy nh bi cc gene nm vng khng tng ng ny ca Y c di truyn theo ng thng (straightforward), ngha l: (i) Chng ch biu hin nam gii; v (ii) chng lun lun c truyn t b cho con trai. 2. S bt hot ca nhim sc th X v mt s vn lin quan

tt c cc ng vt c v k c ngi, nhim sc th X khc bit vi cc nhim sc th khc ch ch c mt chic hot ng trong mt t bo. Cc con ci (gi) c hai nhim sc th X, bnh thng ch mt chic l hot ng v chic kia bt hot. Nhim sc th X bt hot kt vn to thnh mt cu trc bt mu ti sm, thng c dng thu knh li bm vo mt trong mng nhn, gi l th Barr (Barr body) (hnh 4.10). N mang tn ca ngi khm ph u tin l Murray Barr. Cc con c bnh thng ch c mt X v trng thi hot ng trong tt c cc t bo, do vy khng c th Barr. V nguyn tc, s th Barr bng s nhim sc th X ( hnh 4.10B, k hiu l N) tr i mt.
(A) (B) Hnh 4.10 (A) nh chp cc nhn t cc t bo ca (a) mt con ci (XX) vi mt th Barr, (b) mt con c (XY) vi khng th Barr no, v (c) mt con ci XXX vi hai th Barr. (B) S minh ha s hnh thnh th Barr trong cc trng hp khc nhau.
C ch bt hot-X: S bt hot ca mt nhim sc th X cn n mt gene trn nhim sc th ny gi l XIST. Gene XIST m ha mt phn t RNA ln (mt kiu khc vi cc RNA, nh mRNA, c s dng trong tng hp protein). Phn t RNA ca gene XIST tch t dc theo nhim sc th X c cha gene XIST hot ng v gy bt hot tt c (hoc hu nh tt c) hng trm gene khc trn nhim sc th . Mt im cn lu l, RNA ca gene XIST khng di chuyn trn nhim sc th X no khc trong nhn. Cc th Barr l nhng nhim sc th X bt hot c "sn pht " (painted) bi RNA XIST. (Rastan 1994; Lee v Jaenisch 1997). Trong nhng giai on u ca s pht trin phi ci, locus XIST trn mi trong s hai nhim sc th X ca n c biu hin nhng RNA XIST nhanh chng b b gy. Ri mt iu g xy ra thng bo trc s

cn bng i vi mt chic ny hoc chic kia ca cc nhim sc th X. S phin m tip tc trn mt trong s cc nhim sc th X, dn ti vic tch t ca RNA XIST v lm bin di nhim sc th thnh ra mt th Barr bt hot. S phin m ca XIST dng li trn nhim sc th X khc cho php tt c hng trm gene khc ca n c biu hin. S tt ngm ca locus XIST trn nhim sc th X hot ng c tin hnh bng vic methyl ha cc trnh t iu ha XIST (XIST regulator sequences). S methyl ha DNA (DNA methylation) thng xy ra trong biu hin gene cho nn s methyl ha kha cht hn s biu hin ca gene XIST v n cho php biu hin lin tc ca tt c cc gene lin kt-X cn li. Ngoi ra, s bt hot-X phi ci l hon ton ngu nhin i vi mt trong hai X (hnh 4.10B), v n xy ra sm trong qu trnh pht trin phi ( giai on khong 2.000 t bo). Khng th d on liu X t b hay X t m b bt hot trong mt t bo no . Nhng y khng phi l trng hp cho cc mng ngoi phi (extraembryonic membranes); tt c cc t bo ca cc mng ny th ch c nhim sc th X ca ngi b l b bt hot (Rastan 1994; Lee v Jaenisch 1997; Kimball 2004). Mt s gene trn X thot khi s bt hot: Vn t ra l, vi 18 gene c pht hin trn Y cng nh X th sao? S khng c nhu cu no cho cc con ci (females) lm bt hot mt bn sao ca cc gene gi cn bng vi tnh hung cc con c. Th th chng qun l iu tit vic ny ra sao vn cn cha c khm ph (Kimball 2004). Chn on cc bt thng nhim sc th X: R rng l, s hiu bit v hin tng ny c ng dng thc t to ln trong chn on trc sinh hoc kim tra cc bnh tt nhng ngi trng thnh c th c lin quan n cc t bin lch bi nhim sc th X. Chng hn, chn on trc sinh, ngi ta s dng phng php chc i (amniocentesis), ri lm tiu bn kim tra s bt thng nhim sc th v thit lp kiu nhn. Cng thc tng qut cho trng hp ny l: S nhim sc th X = s th Barr + 1 Nh vy, nhng ngi nam bnh thng v cc c th mc hi chng Turner (XO) khng c th Barr no; nhng ngi n bnh thng v cc c th mc hi chng Klinefelter (XXY) c mt th Barr; nhng c th XXX c hai th Barr v.v. (xem Hnh 4.10B). Gii thch s hnh thnh cc th khm di truyn: Vi c ch bt hot ngu nhin ca cc nhim sc th X, cc t bo ca con ci s l th khm di truyn (genetic mosaics) v hai nhim sc th X ca n. y l nguyn nhn dn ti s hnh thnh mu lng "sc vn" hay tam th mo ci (Felix catus). Ta c th hnh dung mi quan h gia kiu gene v kiu

hnh mo lin quan ti mt gene xc nh mu lng trn nhim sc th X nh sau: Nu quy c allele B - mu en v allele O- mu da cam, ta c:
Ci Kiu gene XBXB XXO XBXO
Kiu hnh
Kiu gene
Kiu hnh
en da cam tam th
XBY XOY
en da cam
Nh vy, cho mo c l tam th r rng l n phi c hai nhim sc th X, XBXOY, ngha l th d bi v nhim sc th gii tnh. iu ny rt him khi xy ra trn thc t. n y ta thy rng vic gii thch s xut hin mo tam th bng c ch tng tc gia cc gene allele theo kiu ng tri trong nhiu ti liu trc y v c hin nay l khng ng! cng l l do ti sao Hnh 4.11 S hnh thnh chng ti a mc ny vo y m khng mu sc b lng mo tam th c gii thch bng s t n trong chng 3 (khi xem xt cc th bt hot ngu nhin ca cc t bin lch bi) hoc sau mc II-1 ca nhim sc th X. chng ny nh cch lm truyn thng. 3. Cc tnh trng gii hn bi gii tnh v chu nh hng ca gii tnh Nh ni trn, thng thng cc tnh trng c s phn b khng ng u gia hai gii tnh hoc tp trung mt gii lm ta lin tng ti quy lut di truyn lin kt vi gii tnh do gene hoc trn X (di truyn cho) hoc trn Y (di truyn thng). Tuy nhin, mt s tnh trng c biu hin mt cch khc bit c hai gii nhng khng phi l lin kt vi gii tnh. l nhng tnh trng b gii hn bi gii tnh hoc l chu nh hng ca gii tnh. (1) Cc tnh trng b gii hn bi gii tnh (sex-limited traits): l nhng tnh trng ch biu hin mt gii. C nhiu tnh trng nh vy lin quan vi cc c im sinh sn cc ng vt ci. Chng hn, kh nng cho sa cc gia sc nh tru-b, kh nng trng gia cm v lp chim ni chung, hoc tp tnh trng cc cn trng.
Bng 4.4 V d v tnh trng chu nh hng ca gii tnh Cu HH (Suffolk) Con c Con ci Khng sng Khng sng Hh (Con lai) C sng Khng sng hh (Dorset c sng) C sng C sng

(2) Cc tnh trng chu nh hng ca gii tnh (sex-influenced traits): l nhng tnh trng c xc nh bi cc gene trn nhim sc th thng nhng li biu hin mt cch khc bit hai gii. Chng hn, cc gene xc nh s c sng (horn) cu c biu bin mt cch khc bit cc con c v con ci. mt s ging cu, nh ging cu Suffolk, sng chng thy gii no c (chng c gi l cu khng sng), trong khi cc ging khc, nh ging cu c sng Dorset (Dorset Horn), th c hai gii u c sng mc d cc sng ca con c ln hn sng ca con ci rt nhiu. Khi lai gia hai ging cu ny vi nhau, cc con lai thu c l nhng th d hp t v gene H thuc nhim sc th thng (Hh); tuy kiu gene nh nhau nhng ch c con c mi c sng, cn con ci khng c sng (Bng 4.4). iu d chng t rng, cc c th d hp t, chnh s c mt ca cc hormone c hoc ci xc nh pht trin sng hay l khng pht trin sng. Ngi ta cho rng hin tng hi u sm (premature baldness) ngi, thng xy ra trc tui 35, tun theo cng kiu di truyn nh trng hp va xt cu; ngha l, hi u l tri nhng ngi nam d hp t (Bb) v ln nhng ngi n d hp t (Bb). l l do ti sao chng hi u rt him khi thy ph n.
IV. Lin kt v ti t hp ca cc gen trn mt nhim sc th

Trc khi cp vo cc khm ph ca Morgan v hin tng di truyn lin kt rui gim, ta cn lu rng: Vo nm 1906, Batson v Punnett nghin cu s di truyn ca cc tnh trng khc nhau cy u ngt (sweet pea). Trong mt th nghim kim tra s di truyn ng thi ca mu hoa v hnh dng ht phn, h thu c cc kt qu F2 khng ph hp vi t l 9:3:3:1 ca Mendel. Kt qu cc php lai (k c quy c gene) c trnh by tm tt hnh 4.12. Ptc Hoa ta, ht phn di Hoa , ht phn trn (PPLL) (ppll) Hoa ta, ht phn di F1 (PpLl) F1 (t th phn) F2
Kiu hnh Ta, di Ta, trn , di , trn Tng S quan st (O) 296 19 27 85 427 S k vng (E) 240,2 80,1 80,1 26,7 427,1 (O E)2/ E 13,1 46,5 35,1 127,3 2 = 222,0
Hnh 4.12 Kt qu lai hai tnh ca Bateson v Punnett u ngt.

R rng l t l F2 c quan st v k vng theo t l 9:3:3:1 l hon ton khng khp nhau; nu kim tra bng phng php 2, s sai khc ny cao mt cch ng k v khng ph hp vi gi thuyt phn ly c lp. Hai ng c gng a n v t l 7:1:7:1 gii thch bng kiu tng tc gene, nhng khng thnh cng. V cui cng, h a ra gi rng bi v hai kiu hnh b m vt qu mc cho php F2, c l c mt s ni kt gia cc allele dng b m. i vi hin tng gy nhiu ln s phn ly c lp ny, h gi l kiu kt ni (coupling). Mt khc, h cho rng s lng thp ca cc kiu F2 vn c mt kiu hnh tri gene ny v mt kiu hnh ln gene kia, l do i lc m tnh t nhin ca cc allele tri v ln, v h gi l kiu y nhau (repulsion). 1. Khm ph v s trao i cho rui gim gii thch v mt vt l cc quan st ca Bateson v Punnett, vi nm sau Morgan tin hnh hng lot th nghim s dng ng thi hai gene Drosophila, m mi gene u c hai allele ln v tri (k hiu allele da theo tn ca th t bin ln trong ting Anh nh cp). C th k mt s th nghim m ng lm sau y: (i) mt gene kim sot mu mt (pr: ta v pr+: di) v gene kia kim sot cnh (vg: ngn v vg+: di bnh thng); (ii) mu mt (w: trng v w+: ) v dng cnh (m: cnh b v m+: cnh bnh thng); hoc (iii) mu thn (e: mun v e+: xm) v dng cnh (c: cnh cong v c+: cnh thng) v.v... By gi ta hy xt mt trng hp quen thuc, l: gene quy nh mu sc thn (allele ln b: en v allele tri B: xm) v gen xc nh chiu di cnh (vg: cnh ngn v Vg: cnh di bnh thng). Morgan cho lai gia hai dng rui thun chng thn xm, cnh di (BBVgVg) v thn en, cnh ngn (bbvgvg), i con F1 ng thu c tt c cc con c v ci u c thn xm, cnh di (BbVgvg). Nhng khi em lai phn tch cc con c v ci F1, kt qu hon ton khc nhau. Trc tin, ta xt php lai phn tch gia cc con ci xm-di F1 vi con c en-ngn. Kt qu cc php lai ny cho i con c c bn kiu hnh (xem hnh 4.13a), nhng vi t l quan st khc xa vi t l k vng 1:1:1:1 (xem hnh 4.13b), nh sau:
Kiu hnh xm-di en-ngn xm-ngn en-di Tng
S liu quan st S liu k vng
965 575
944 575
206 575
185 575
2300 2300
Cc kiu hnh b m
Cc kiu hnh ti t hp
T kt qu i con ca lai phn tch rui ci F1 cn cho thy cc kiu hnh b m ban u l xp x nhau, chim 83% ([965 + 944]: 2300 =

0,83); cc kiu hnh ti t hp cng c s lng gn nh tng ng, chim 17% ([206 + 185]: 2300 = 100% 83% = 0,17). iu ny ch c th l gii nu nh cc gene nm trn cng nhim sc th, hay gi l lin kt (linkage), v trong qu trnh gim phn rui ci c xy ra s trao i cho (crosing over) hay ti t hp (reconbination) gia hai gene ny. Tht vy, trong php lai phn tch ny, rui c en-ngn (bvg/bvg) ch cho mt loi giao t cha c hai allele ln, bvg). T y suy ra rui ci F1 xy ra trao i cho v cho bn loi giao t tng ng vi t l cc kiu hnh i con: (i) Cc kiu b m (parental) mi kiu c t l trung bnh l 41,5% (BVg = bvg = 0,415); v (ii) Cc kiu ti t hp (recombinant) mi kiu c t l trung bnh l 8,5% (Bvg = bVg = 0,085). Nh th, da vo thnh phn gene ca P hoc thnh phn allele ca cc giao t chim t l cao (tc cc kiu b m) ta d dng xc nh c kiu gene ca F1 l BVg/bvg (kiu cis).
Kiu di en, ngn
Kiu gene b m
Kiu gene ti t hp
en-ngn
Kiu di
Xm-ngn
en-di
Kiu hnh b m
Kiu hnh ti t hp
Hnh 4.13 Kt qu ca cc php lai P v lai phn tch rui ci F1 (tri) v gii thch bng c s t bo hc ca kt qu lai phn tch rui ci F1.
Trong th nghim th hai, Morgan lai hai kiu gene, mi kiu l ng hp v allele tri mt gene ny v ng hp v allele ln mt gene kia (BBvgvg bbVgVg). Cc rui F1 d hp t kp (BbVgvg) sau c lai vi dng kim tra bbvgvg. Kt qu ca php lai ny, so vi th nghim trc, cho thy i con c s o ngc gia nhm cc kiu b m v nhm cc kiu ti t hp; ngha l, s lng i con mang cc tnh trng tri n (Bvg/ bvg v bVg/ bvg) cao hn nhiu so vi k vng, trong khi s lng i con mang cc tnh trng tri kp (BVg/bvg) v cc tnh trng ln kp (bvg/ bvg) thp hn nhiu so vi k vng. Nh vy, F1 c kiu gene Bvg/bVg (kiu trans).

Ghi nh: (1) S kin trao i cho thng c ch ra trn cc s bng mt 'du cho' () ni gia cc nhim sc th tng ng (hnh 4.14). (2) i vi trng hp d hp t kp v hai cp gen lin kt, c hai AB Ab cch sp xp cc allele trn mt cp nhim sc th: v , m c th ab aB vit n gin l: AB/ab v Ab/aB. - Cch sp xp cc allele trong hai allele tri trn mt nhim sc th v hai allele ln trn chic kia, AB/ab, c gi l kiu kt ni (coupling) hay kiu u (cis). Vi kiu gene ny, AB v ab l cc giao t thuc kiu cha m, cn Ab v aB l cc giao t thuc kiu ti t hp. - Cch sp xp cc allele trong mi nhim sc th mang mt allele tri v mt allele ln sao cho trong th d hp kp chng cc v tr cho nhau, Ab/aB, c gi l kiu y nhau (repulsion) hay kiu lch (trans). Vi kiu gene ny, Ab v aB l cc giao t thuc kiu cha m, cn AB v ab l cc giao t thuc kiu ti t hp.
Hnh 4.14 M hnh mt trao i cho n (tri) v cch biu din n.
(3) S ti t hp ca cc gene lin kt kiu u (cis) cng nh kiu cho (trans) t c hai php lai phn tch ni trn v cc php lai thun nghch ni chung u xy ra vi tn s gn nh nhau; (4) Tng t l ca cc kiu ti t hp i con ca php lai phn tch c gi l tn s ti t hp, v tn s ny c trng cho mi cp gene ring bit (nh s tho lun di y). 2. Lin kt gene hon ton (hay gim phn khng c trao i cho) By gi ta tr li th nghim u tin ca Morgan. Khi thc hin php lai phn tch gia cc con c xm-di F1 vi con ci en-ngn, i con ch c hai kiu hnh xm-di v en-ngn vi t l xp x 1:1, ch khng phi bn kiu vi t l u nhau nh d on. iu ny chng t rui c F1 ch cho hai loi giao t vi t l tng ng ( BVg = bvg = 50%), tng ng vi hai kiu hnh i con (v rui ci en-ngn ch cho mt loi giao t cha hai allele ln, bvg); ngha l cc gene ny cng nm trn mt nhim sc th v gia chng c s lin kt hon ton (complete linkage). Ni cch khc, ti t hp khng xy ra rui gim c.

Gim phn m khng c bt k s trao i cho no ch xy ra mt s t loi. cc sinh vt ny, ti k gia I cc nhim sc th tng ng nm dc bn nhau mt phng ca thoi. Chng phn tch theo cch thng thng k sau I, v gim phn tin hnh mt cch bnh thng sau . Kiu gim phn ny thy c mt vi cn trng, k c cc con c ca rui gim, v mt s thc vt c hoa. tt c cc loi cn li c t nht mt trao i cho c hnh thnh trong mi th lng tr k trc I. Tm li, tt c cc loi sinh sn hu tnh, trong qu trnh gim phn c th khng xy ra trao i cho hoc c t nht mt trao i cho c hnh thnh trong mi th lng tr k trc I. i vi hai cp gene bt k trn mt cp nhim sc th tng ng, cc sn phm tng ng c to thnh trong cc giao t c minh ha hnh 4.15 di y.
A. Khng trao i cho
B. C trao i cho
Hnh 4.15 Gim phn khng c trao i cho (A) v c mt trao i cho n (B), v cc sn phm c to thnh i vi hai cp gene bt k trn mt cp nhim sc th tng ng.
V. Trao i cho v lp bn di truyn

1. Tn s ti t hp S trao i cho l mt qu trnh trao i gia cc nhim sc th trong gim phn (kt hp vi s hnh thnh giao t mt cch bnh thng) cho ra cc t hp tnh trng mi, gi l cc bin d t hp (xem hnh 4.13). Tn s ti t hp (frequency of recombination or rate of recombination), cn c gi l tr s trao i cho (cross over value), k hiu l r, c tnh bng t l phn trm ca cc th ti t hp sinh ra trong mt php lai phn tch, theo cng thc sau: n r = 100 m

trong : n - s lng cc c th ti t hp c sinh ra, v m - tng s c th ca i con ca php lai phn tch; v r - tn s ti t hp (thng gi l tn s hon v gene), l mt s hu t tha mn min gii hn [0; 0,5], c th biu din bng s thp phn hoc phn trm. cc sinh vt m ch s trao i cho c nghin cu rng ri, chng hn nh ng v rui gim, t l ca cc th ti t hp sinh ra trong mt php lai c th l kh n nh. Tuy nhin, cc t l ny l khc nhau i vi cc gene khc nhau trn cng mt nhim sc th. iu ny c gii thch da trn c s rng mi mt gene c mt v tr c nh (tc locus ca n) trn mt nhim sc th c th, v rng s trao i cho c th xy ra gia cc gene nm xa nhau. Vi v d v hai gene b v vg rui gim ni trn, ta c th ni rng cc locus ny nm kh gn nhau, bi v c rt t cc th ti t hp c to ra. p dng cng thc tnh tn s ti t 206 + 185 hp trn, ta tnh c: r = 100 = 17% . T y suy ra t l 2300 k vng ca cc loi giao t ti t hp v khng ti t hp (dng b m): giao t kiu ti t hp Bvg = bVg = 17%: 2 = 8,5% BVg = bvg = (100% 17%): 2 = 41,5% giao t kiu b m Mt cch tng qut, da theo tn s ti t hp (r), ta c th biu din t l ca cc loi giao t ti t hp v khng ti t hp i vi hai kiu gene d hp t u (AB/ab) v d hp t cho (Ab/aB) bng 4.5.
Bng 4.5 T l ca cc loi giao t Giao t AB ab Ab aB Tng Kiu gene b m AB/ab (1 r) (1 r) r r 1 Ab/aB r r (1 r) (1 r) 1
By gi ta cp mt t v mi quan h gia hai gene khng allele trong qu trnh gim phn to giao t, m ta c th kim tra bng phng php kinh in c p dng trong nghin cu di truyn hc l lai phn tch. Ngoi ra, cng c th s dng cc phng php khc, khng c ph bin lm hoc kh phc tp nh l t th phn hoc tp giao. i vi hai gene trng thi d hp t kp quy nh hai cp tnh trng tng phn khc nhau, trong mt php lai phn tch:

(i) Nu nh hai gene nm trn hai nhim sc th khc nhau, s c bn kiu giao t (hay kiu hnh) c to thnh vi t l ngang nhau; (ii) Nu nh hai gene lin kt hon ton trn mt nhim sc th, ch c th to ra hai loi giao t (hay kiu hnh) vi t l ngang nhau (r = 0); (iii) Nu nh hai gene lin kt khng hon ton trn mt nhim sc th, c th to ra bn loi giao t (hay kiu hnh) thuc hai nhm, trong cc kiu giao t ti t hp mi kiu chim t l bng r/2, v cc kiu giao t b m mi kiu chim t l bng (1 r)/2; (iv) Tn s ti t hp (r) bin thin trong khong 0 0,5; thng thng r thp hn 50%, ph thuc vo khong cch vt l gia hai gene trn nhim sc th. Trong cc trng hp cc oan, tr s r c th bng zero (r = 0), nu nh hai gene lin kt hon ton hoc nm gn st nhau; v tr s r ny cng c th t cc i, ngha l r = 0,5 hay 50%, trong trng hp cc gene nm cch nhau hn 50 n v bn (nh chng ta s cp ngay di y) hay ni cch khc, tt c cc t bo sinh giao t tri qua gim phn u xy ra trao i cho gia hai gene c xt n. Khi t l cc giao t (hay t l phn ly kiu hnh) trng vi trng hp cc gene phn ly c lp. Tuy nhin, rt him khi xy ra nh vy. 2. Bn di truyn Vic thit lp bn gene ca cc nhim sc th ln u tin c p dng rui gim Drosophila, c xut vo nm 1913 bi Alfred Sturtevant, mt mn ca Morgan. Theo ng, ta c th da vo cc tn s ti t hp ca cc gene thu c trong cc php lai phn tch m t mi quan h vt l ca cc gene trn mt nhim sc th theo trt t tuyn tnh, gi l bn lin kt (linkage map) hay bn di truyn (genetic map). Khong cch bn (map distance) gia hai gene, v d b v vg l 17%, c coi l cch nhau 17 n v bn (map unit; vit tt l: m.u.); hay ni cch khc, 1 n v bn l 1% ti t hp. Cc nghin cu v sau cho thy rng khong cch di truyn c o bng phng php thng k (tc t l phn trm ti t hp, do Morgan v Sturtevant xut) ni chung l ging vi cc khong cch trn nhim sc th o c v mt t bo hc hay ha sinh hc. Nguyn tc chung ca vic xy dng bn nhim sc th mt loi no , theo Morgan v cc ng s ca ng, c th tm tt th ny: (1) Xc lp s nhm lin kt (s nhim sc th n bi) ca loi. iu ny c th tin hnh bng cch thc hin hng lot cc php lai c th c xc nh cc mi quan h (c lp v lin kt) trong s hng lot gene, kt hp vi cc nghin cu t bo hc (m s lng nhim sc th).

(2) Xc nh thnh phn gene ca mi mt nhm lin kt (da vo cc kt qu lai khc nhau). (3) Xc nh v tr, trt t v khong cch gia cc gene trn mi nhm lin kt. Nguyn tc chung l, gia cc gene lin kt c tn s ti t hp cng thp chng no chng t chng nm cng gn nhau chng y; theo chui suy lun ta c th bit c v tr ca mt gene khi u v cc gene k tip theo chiu dc trn mt nhim sc th, v t gene khi u nm u mt ca vai ngn ng vi v tr zero (0,0); sau dng phng php cng dn biu din v tr v khong cch trn bn ca cc gene k tip vi so vi gene u mt ny. Khi ta c c bn ca mt nhm lin kt. Ta c th hnh dung n nh mt on thng nm ngang c nhiu vch ch ra trt t thng hng ca cc gene, mi vch ng vi mt locus-gene c th (pha trn vch l k hiu v c th c tn y ca th t bin gene v pha di vch l v tr trn bn ca gene so vi gene khi u, c ghi bng mt con s c th; hnh 4.17a). Nu ta dng ng bn ln, theo nguyn tc, k hiu cc gen s nm v pha bn phi v khong cch bn ca cc gene nm pha bn tri (hnh 4.17b). y, c mt s im cn lu : (i) chnh xc ca bt k bn lin kt no c thit lp trc y u c th thay i v ngy cng chnh xc hn; l do cc pht hin b sung nhng gene mi nm trung gian gia cc gene c nh v cng vi cc tn s ti t hp ca chng; (ii) Ring rui gim, nhim sc th X c Morgan gn cho s I, k n l cc s II, III v IV tng ng vi s nh dn v kch thc. Ngy nay, mc d h thng nh s nhim sc th c thay i, song i vi rui gim vn c gi nguyn biu th lng tn knh i vi Morgan - ngi sng lp thuyt di truyn nhim sc th; v (3) Morgan ci tin cch lp bn vn cng knh phc tp ni trn bng phng php lai phn tch ba im (nh s c tho lun di y).
Bng 4.6 Tn s ti t hp ca nm cp gene lin kt-X Cc gen thn vng (y) mt trng (w) thn vng (y) mt ti (v) mt trng (w) mt ti (v) mt ti (v) cnh b (m) mt trng (w) cnh b (m) mt trng (w) cnh khng pht trin (r) mt ti (v) cnh khng pht trin (r) Tn s ti t hp 214/21.736 = 0,010 1.464/4.551= 0,322 471/1.584 = 0,297 17/573 = 0,030 2.062/6.116 = 0,337 406/898 109/405 = 0,452 = 0,269

Chng hn, bng cch s dng s liu bng 4.6, Sturtevant xc nh cc mi quan h vt l ca nm gene ny v gi rng chng xp theo mt ng thng. Cch lm nh sau: (1) Hai gene y v w c tn s nh nht (ry-w = 1%) nn nm gn nhau. (2) v rw-v = 29,7% < ry-v = 32,2% trt t ba gene ny phi l y-w-v. (3) Gene gn nht vi v l m (rv-m = 3%), m rw-m rw-v + rv-m m phi nm bn phi v. (4) rv-r = 26,9% < rw-r = 45,2% r phi nm bn phi v v m. Vy, trt t ca nm gene lin kt trn X l: y-w-v-m-r. T y, Sturtevant xy dng nn mt bn bt u bng y v tr 0,0 v pha bn tri v s dng cc tn s ti t hp gia cc gene k nhau biu th v tr ca chng trn bn ng vi tng con s c th. Theo , w v tr l 1,0, v v tr 30,7 (= 1,0 + 29,7), m v tr 33,7 (= 30,7 + 3,0), v r v tr 57,6 (= 30,7 + 26,9). So vi bn hin nay, v tr nm gene trn bn ca Sturtevant c hi khc mt cht bi v c nhiu gene trung gian c pht hin b sung (hnh 4.16).
yw Sturtevant 0,0 1,0 Hin nay 0,0 1,5 m 30,7 33,7 33,0 36,1 v r 57,6 54,5 Tm ng 66,7
Hnh 4.16 Bn Sturtevant v nm gene lin kt-X v so vi hin nay.
Di y cho thy bn ca nhim sc th s 2 rui gim (hnh 4.17a) v nhim sc th s 9 ng (hnh 4.17b) vi nhng gene chnh yu. Chng hn, trn hnh 4.17a, hnh ng gia tng trng cho nhim sc th s 2 rui gim ch ra v tr ca su gene; pha trn l cc kiu hnh t bin, v pha di l cc kiu hnh bnh thng. T tri sang: v tr 0 gene al (aristaless antenna) ch t bin ru ngn i vi ru di bnh thng; v tr 13 - gene dp (dumpy wings) ch t bin cnh xn vi cnh di; v tr 31 - gene d (dachs) ch t bin chn ngn vi chn di), v tr 54,5 - gene pr (purple eyes) ch t bin mt ta vi mt ), v tr 67,0 - gene vg (vestigial) ch t bin cnh ngn vi cnh di), v tr 104 - gene bw (brown eyes) ch t bin mt nu vi mt ). Cn lu rng, ngy nay, sau khi nh v bn cc gene xc nh by tnh trng m Mendel nghin cu u H Lan, ngi ta khng khi ngc nhin ti sao Mendel li khng quan st hin tng lin kt trong cc php lai hai tnh ca mnh. Mc d Mendel chn ra by tnh trng v c by nhim sc th u H Lan (2n = 14), hai trong s cc tnh trng ng nghin cu c xc nh bi cc gene trn nhim sc th s 1 v ba tnh trng c xc nh bi cc gene trn nhim sc th s 4 (bng 4.7). Thc ra, Mendel ch chn nghin cu mt vi tnh trng v c l, tht may mn l ng khng quan st thy cc kiu m khc vi k

vng v s phn ly c lp. Chnh iu ny lm nn s nht qun hon ton v ht sc r rng, n gin trong cc kt qu m ng cng b.
Hnh 4.17 Bn nhim sc th s 2 ca rui gim (bn tri) v nhim sc th s 9 ca ng (bn phi).
Qu ng nh Watson vit trong Li ni u cun "Chui xon kp - Hi k v vic pht minh ra cu trc ca DNA" ca mnh, rng: "Cch gii quyt ng khng nhng phi p m cn phi n gin" (Watson 1968). Chn l bao gi cng gin d nhng gin d cha hn l chn l; chn l bao gi cng p nhng p th cha l chn l. Tuy nhin, nhn chn c ci gin d, ci p ch thc trong cc khm ph ca Mendel, nh chng ta u bit, nhn loi phi mt chng 35 nm!
Bng 4.7 Cc v tr nhim sc th i vi by tnh trng u H Lan Tnh trng Dng ht Mu sc ht Mu sc qu Kt cu qu Mu sc hoa V tr hoa Chiu cao cy Kiu hnh trn-nhn vng-xanh xanh-vng trn-nhn tm-trng trc-nh cao-thp Allele R-r I-i Gp-gp V-v A-a Fa-fa Le-le Nhim sc th 7 1 5 4 1 4 4

3. Trao i cho bn si Hc thuyt ca Morgan v cc gene lin kt v trao i cho c ng h rng ri bi nhiu s liu. Tuy nhin, cn cha tht s r rng ch, liu s ti t hp xy ra gia cc nhim sc th trc khi chng ti bn ( giai an hai si) hay sau ti bn ( giai on bn si). Nu nh trao i cho xy ra trc khi ti bn th tt c bn chromatid s l cc th ti t hp, trong khi nu xy ra sau ti bn th ch c hai trong bn chromatid s l cc th ti t hp (xem hnh 4.15). phn bit gia cc dng bin i ny, phi cn ti mt h thng di truyn trong tt c cc sn phm di truyn t gim phn phi c xc nh. Bng chng u tin v s trao i cho sau ti bn nhim sc th c c t vic nghin cu rui gim, trong hai nhim sc th X ca rui gim ci dnh nhau tm ng ca n v to thnh mt nhim sc th sai hnh, gi l nhim sc th X-dnh (attached-X). Kt qu ca s dnh ny l, t con ci c nhim sc th X-dnh v mt nhim sc th Y tch ring (X-XY) s cho ra cc giao t hoc cha hai nhim sc th X hoc khng c X (ch c mt Y). V d, php lai trong c hai nhim sc th X u c mang allele kiu di ti locus mt trng c cho hnh 4.18. Cc con ci X-XY vi allele kiu di trn c hai X (X+-X+Y) c cho lai vi cc con c mt trng "bnh thng" (XwY). Cc c th con sng c u ging vi b m, trong khi cc con rui gim YY cht ht v XXX thng l b cht.
X+-X+Y XwY (ci mt ) (c mt trng) + + (Xw : Y) Giao t (X -X : Y) : X+-X+Y : XwY : YY F1 Kiu gene = X+-X+Xw Kiu hnh = (thng cht) : (ci mt ) : (c mt trng) : (cht)
Hnh 4.18 Mt php lai gia rui gim ci kiu di X-dnh v con c mt trng. y i con sng st th ging ht cc b m chng.
Khi cc allele trn hai nhim sc th X-dnh l khc nhau th s trao i cho hai si v bn si l c th phn bit c. Chng hn, hnh 4.19 cho thy th t bin mt thi Bar lm gim s lng cc mt b t con s bnh thng khong 780 kiu di (con ci B+B+ hoc con c B+Y) xung cn khong 360 cc th d hp (con ci BB+) v khong 70 cc th ng hp (con ci BB hoc con c BY). Bng cc thc nghim cho thy rng: (i) Trong trng hp khng xy ra trao i cho, tt c cc giao t mang cc nhim sc th X th c dng

X-dnh trng thi d hp v do c kiu hnh mt Bar d hp. (ii) Nu trao i cho xy ra giai on hai si th s c dnh lu ti c hai si. V vy, khi ti bn xy ra sau , tt c cc giao t s li l d hp. Ni cch khc, cc kt qu quan st c trong c hai trng hp khng c trao i cho v trao i cho hai si l nh nhau.
(a)
(b)
Hnh 4.19 (a) Cc kiu hnh di, mt Bar d hp v ng hp (ultraBar); v (b) Cc kiu bng tng ng c mt, hai v ba bn sao ca bng 16A.
Ti t hp giai on bn si sinh ra cc kt qu khc nhau i con. Trong trng hp ny, ch c hai trong bn si tham gia vo ti t hp, v vy s sinh ra cc kiu mi. Vi ti t hp giai an bn si, c kiu di ln kiu ng hp BB s c to ra. Trn thc t, iu ny c quan st trong cc th nghim, ch ra rng s ti t hp xy ra sau khi ti bn ngha l, trao i cho phi xy ra gai on bn si (xem hnh 4.15). Chng ta s cp tr li bng chng b sung v trao i cho bn si vi nm Neurospora trong phn sau (mc VII). S lc v trao i cho phc (multiple crossing over) Trn thc t, khi hai gene nm xa nhau trn mt nhim sc th, gia chng c th xy ra nhiu trao i cho trong mt ln gim phn v iu ny gy ra nhng rc ri trong vic tnh ton, gii thch cc s liu ti t hp v lp bn da trn cc s liu ny. Trn hnh 4.20 cho thy trao i cho kp giai on bn si c th xy ra theo nhiu cch khc nhau. Chng hn, hnh 4.20a cho thy trao i cho kp xy ra gia hai cp gene Aa v Bb nhng cc nhim sc th ny khng ti t hp i vi cc allele ca hai gene, v do khng th phn bit c vi cc nhim sc th khng trao i cho v mt di truyn. Kt qu l cho ra ch hai loi giao t vi t l ngang nhau (AB = ab) ging nh trng hp khng trao i cho

(hnh 4.15A), thay v cho bn loi giao t thuc hai kiu (hnh 4.15B). Hin tng ny lm cho gi tr ti t hp nhn c s thp hn tn s trao i cho thc v nh hng ti khong cch bn gia cc gene.
(a)
(b)
Hnh 4.20 (a) Hai trao i cho n xy ra vng gia hai gene A v B, v kt qu l sinh ra c bn loi giao t khng ti t hp i vi hai gene ny, ngha l khng pht hin c th trao i cho kp no. (b) Trao i cho n v cc dng trao i cho kp lin quan n hai, ba v c bn si c th xy ra trong mt cp nhim sc th tng ng.
Nh ni, tn s ti t hp cc i gia hai gene bt k trn mt cp nhim sc th tng ng l 50%. iu ny ch c th xy ra khi hai gene nm cch xa nhau ti mc t nht c mt on trao i cho hu nh lc no cng xy ra gia chng. Trn hnh 4.15B ta thy r rng, c mt trao i cho n trong mi ln gim phn s cho ra mt na s sn phm c cc t hp ging b m v na kia l cc sn phm c ti t hp gene. Trn hnh 4.20b cng cho thy khi xy ra hai trao i cho n m c mt chromatid chung hay trao i cho kp lin quan ba si (hai hnh u ca hng di), th kt qu s khng khc vi kt qu ca trao i cho n nh hnh 4.15B. Mt kh nng khc, l trao i cho kp lin quan c bn si (hnh 4.20b, xem hnh cui ca hng di), trong s trao i cho th hai xy ra gia hai chromatid ( ngoi) khc vi hai chromatid tham gia vo trao i cho th nht ( trong). Trong trng hp ny, tt c cc sn phm u c ti t hp. Nu cc chromatid tham gia vo hai trao i cho n mt cch ngu nhin th t l k vng ca ba kiu trao i cho kp s tng ng vi t l : : . iu c ngha l trung bnh s c hai trong bn sn phm ca gim phn l c ti t hp:

()(0) + ()(2) + ()(4) = 2. T l ny r rng l ging nh khi mt trao i cho n m lc no cng xy ra gia hai gene. Thc ra, nh s cp di y, trao i cho kp c th pht hin c trong cc th nghim ti t hp s dng ba gene, gi l lai ba im.
VI. Lp bn gene t cc php lai phn tch ba im

1. Trao i cho kp vi vic xc nh trt t v khong cch cc gene Gi s ta c ba gene lin kt A, B v C cn lp bn . Theo l thuyt, c th c ba cch sp xp khc nhau ty vo v tr gene gia, l: (i) A-B-C; (ii) A-C-B; v (iii) B-A-C. Tuy nhin, trong trng hp cha bit chc v s lin kt ca ba gene, v gi s ta xc nh c tn s ti t hp gia hai gene A v B l 15% v gia hai gene B v C l 20%. Theo suy lun, ta bit rng ba gene ny cng nhm lin kt; v theo nguyn tc, chng c th lin kt vi nhau theo mt trong hai cch: (i) A-B-C; hoc (ii) B-A-C. Trong trng hp ny, ta cn phi tin hnh mt php lai phn tch gia A v C xem liu tn s ti t hp l 35% hay 5%. Nh vy, xc nh c v tr tng i ca ba gene ta phi tin hnh ba php lai phn tch ring bit. Trong khi , nu da vo kh nng xy ra mt trao i cho kp hay l hai trao i cho n ng thi (trong trng hp ba gene khong cch ln), ta ch cn thc hin mt php lai phn tch ba im (three-point testcross) l . iu c ngha l, vic lp bn ba gene tr nn gn nh, khng cn cng knh phc tp nh trc na.
(a)
(b)
Hnh 4.21 (a) Mt trao i cho kp, v (b) cc sn phm to thnh.
Tht vy, theo l thuyt xc sut ca cc s kin ngu nhin, mt khi trao i cho kp xy ra ( mt s t bo ny), th cc trao i cho n ring r (d nhin l xy ra cc nhm t bo khc) tt yu phi xy ra v c cc tn s cao hn hn tn s trao i cho kp. Trong trng hp , i con ca php lai phn tch thu c (nu y ) s c tm kiu hnh thuc bn nhm khc nhau: Trc ht l cc kiu cha m vi tn s cao nht; k n l cc kiu trao i cho n khc nhau; v cui cng l cc kiu trao i cho kp vi tn s thp nht. V theo nguyn tc, c hai kiu hnh ca mi nhm s c s lng tng ng nhau. Trn hnh 4.21 cho thy mt trao i cho kp xy ra on cha gene b (nm gia a v

c), v kt qu l cho ra bn sn phm gm hai kiu cha m (abc v + + +) v hai kiu ti t hp i vi gene b (a + c v + b +). Vi kt qu ny ta d dng xc nh gene gia v n l sn phm ca trao i cho kp. By gi ta hy tm hiu hai v d kinh in v lai phn tch ba im, mt th nghim rui gim v mt th nghim chn ging ng. V d 1: Trong th nghim s dng li ba gene lin kt-X rui gim (y, w v m), sau khi thu c cc con ci F1 d hp t v ba cp gen, Morgan cho chng lai tr li vi con c dng kim tra (hnh 4.22). Kt qu phn tch cho thy cc con ci F1 cho tm loi giao t (tc tm kiu hnh i con ca n), hai trong s l cc kiu giao t b m v su kiu cn li l cc giao t ti t hp. Theo s trnh by trn, ta c th tin hnh phn loi v tnh tn s ca mi kiu giao t da vo s lng cc kiu hnh thng k c (hnh 4.22) ri v bn cho ba gene ny, theo cc bc chung nht nh sau: P +++/+++ ywm/Y +++/ywm ( F1) ywm/Y ( dng kim tra)
S lng 3.501 3.471 1.754 1.700 28 32 6 3 10.495 Tn s 0,664 0,329 0,0057 0,00086 1,0
Kiu b m (khng ti t hp) Kiu ti t hp n m vi w Kiu ti t hp n y vi w Kiu ti t hp kp w vi y v m Tng
Giao t +++ ywm ++m yw+ y++ +wm +w+ y+m
Hnh 4.22 Mt php lai phn tch cc rui gim ci d hp t v ba gene lin kt -X (c cho sn s lng v tn s ca cc kiu khc nhau i con).
Bc 1: Xc nh thnh phn gene tri ln trong nhm lin kt (da vo cc kiu giao t b m, tc hai kiu hnh c s lng nhiu nht). y l y w m v + + + ( bit trc). Bc 2: Xc nh v tr gene gia. Cn c vo hai kiu hnh c s

lng t nht suy ra y l sn phm ca trao i cho kp (Lu : trn thc t, c th ch xut hin mt kiu hnh c th l do s lng cha ln hoc do cc gene nm kh gn nhau nn gy nhiu, nh s tho lun phn sau). Qua so snh thnh phn gene ca cc kiu ti t hp kp vi cc kiu b m cho php tm ra gene thay i v tr chnh l gene gia. Trong v d ny, gene gia l w. Vy trt t ba gene l y-w-m, v kiu gene ca con ci F1 l: + + + / y w m. Khi bit c gene gia, ta d dng xc nh c cc kiu trao i cho n bng cch sp xp li trt t gene cc kiu giao t ri so snh vi cc kiu b m tm ra gene c s thay i v tr. V d, vi hai kiu giao t + + m v y w +, v m i ch nn y l kt qu ca trao i cho n gia w v m. Vy hai kiu cn li l do trao i cho n gia y v w. Bc 3: Tnh khong cch gia cc gene. tm khong cch gia cc gene c th tin hnh theo mt trong hai cch sau: (i) Tnh trc tip da vo s lng ca cc kiu ti t hp. Theo nguyn tc, khong cch ca hai gene nm k nhau th bng s lng c th ca cc kiu ti t hp gia hai gene cng vi s lng c th ca cc kiu ti t hp kp ri chia cho tng s c th i con. V khong cch gia hai gene u mt th bng tng cc khong cch ca cc gene nm trong n. C th, khong cch y v w = (28 + 32 + 6 = 3) / 10.495 = 0,0066. Tng t, ta tnh c khong cch hai gene w v m = (1.754 + 1.700 + 6 = 3) / 10.495 = 0,330. Khi khong cch bn gia y v m = 0,0066 + 0,330 = 0,3366. (ii) Tnh gin tip thng qua cc tn s kiu hnh thuc cc nhm khc nhau (xem hnh 4.22). Khi c c cc tn s ny ri, ta tnh khong cch gia hai gene k nhau bng cch cng tn s trao i cho n thc t ca hai gene vi tn s trao i cho kp thc t. V d, khong cch hai gene y v w = 0,0057 + 0,00086 = 0,0066; v khong cch hai gene w v m = 0,329 + 0,00086 = 0,330. Khi khong cch y v m l 0,3366 (Cng c th tnh khong cch y v m bng cch cng hai tn s trao i cho n thc t; ngha l bng 0,0057 + 0,329 = 0,335. Con s ny c hi khc mt cht so vi con s tnh c trn. Tuy nhin, trn thc t, vic tnh khong cch hai gene u mt l khng cn thit). Bc 4: V bn gene. Bn hy t v bn cho ba gene ny. V d 2: Tm tt kt qu th nghim lai phn tch ba gene trn trn mt nhim sc th ng (Zea mays). Cc allele ln c s dng trong php lai mt c th F1 d hp t ba cp gene (Lz Gl Su / lz gl su) ny l: lz (tp tnh mc b lan), gl (l bng), v su (ni nh ng). Php lai v kt qu c cho bng 4.8.
(F1 em lai) Kiu hnh i con ca php lai phn tch bnh thng (kiu di) mc b lan l bng ni nh ng mc b lan, l bng mc b lan, ni nh ng l bng, ni nh ng mc b lan, l bng, ni nh ng
(dng kim tra)
Bng 4.8 Kt qu i con ca php lai phn tch ba im ng Kiu gene giao t t F1 Lz Gl Su lz Gl Su Lz gl Su Lz Gl su lz gl Su lz Gl su Lz gl su lz gl su Tng S lng 286 33 59 4 2 44 40 272 740
Bng phng php trn y, ta xc nh c: (i) Thnh phn nhm gene lin kt phi l: Lz Gl Su ( bit trc) (ii) Gene gia l Su, v trt t ba gene: Lz-Su-Gl (iii) Khong cch gia Lz v Su = (40 + 33 + 4 + 2)/ 740 = 0,107 hay 10,7%; gia Su v Gl = (59 + 44 + 4 + 2)/ 740 = 0,148 hay 14,8%. (iv) Bn gene v s trao i cho kp c minh ha nh sau:
2. nhiu (interference) v h s trng hp (coefficient of coincidence) V thc cht, ti t hp hay trao i cho l s kin mang tnh xc sut cho nn iu quan trng cn bit l xem liu cc trao i cho n ny l c lp vi nhau hay c nh hng no ln nhau hay khng. Nu cc s kin ti trao i cho khc nhau l c lp, th xc sut ca hai s kin ng thi nh th xy ra trong cng giao t s bng tch xc sut ca cc s kin ring r. Khi , tn s trao i cho kp k vng t v d 1 s bng 0,330 0,0066 = 0,00218. Trong khi , tn s trao i cho kp

quan st c l 0,00086; r rng l thp hn t l c k vng. ng ra, theo l thuyt, s c th trao i cho kp xut hin phi bng 0,00218 10495 23 (con s thc t xut hin t hn 23 9 = 14 c th). S chnh lch nh th ni chung rt ph bin cc sinh vt, v n c coi l hin tng nhiu (interference); ngha l s xut hin mt trao i cho mt vng nhim sc th lm gim xc sut xy ra trao i cho vng th hai ln cn n. m t s khc nhau gia cc s lng trao i cho quan st v k vng , ln u tin H.J. Muller a ra mt phng php chun cho s tnh ton ny. Theo , h s trng hp (coefficient of coincidence) c o bng s lng hay tn s cc th ti t hp kp quan st (O: observed) chia cho s lng hay tn s k vng tng ng (E: expected); v nu ta k hiu h s trng hp l C, th C = O/E; trong : 0 C 1. Tr s ny l mt tiu chun cho php o mc nhiu I (interference) ca mt trao i cho ny ln mt trao i cho khc. Cng thc dng tnh nhiu ny nh sau: I=1C trong 0 I 1. T v d 1 trn, ta tnh c C = 0,00086/0,00218 = 0,3945; iu ny c ngha l s lng trao i cho kp thc t xy ra ch bng 39,45% con s l thuyt. Vy I = 1 0,3945 = 0,6055. Tr s ny cho thy mc nhiu cao ng k. R rng hai i lng ny l nghch nhau v tng ca chng bng n v (I + C = 1). Nu I = 1, lc C = 0; iu c ngha l, hin tng nhiu tng dn ln khi m khong cch hai gene pha bn ngoi cng ngn li, cho ti khi trao i cho kp khng th xy ra ti mt im gii hn (nhiu hon ton) v h s ph hp bng zero. nhiu sinh vt khong cch ny l khong 10 n v bn . Ngc li, nu I = 0, th C = 1; ngha l khi cc gene nm rt xa nhau n ni cc trao i cho ca chng khng nh hng g ln nhau (hon ton khng nhiu), th hin tng nhiu bin mt v h s ph hp bng 1. Cc thc nghim cho thy I = 0 khi tng khong cch gia cc gene ln hn chng 45 n v bn . Mt l thuyt gii thch tn s trao i cho kp thp him l ch tr s nhiu cao i vi cc gene lin kt cht r rng l c lin quan vi hin tng "x cng", khng th vn xon v mt vt l ca cc chromatid trong mt vng rt ngn. Tuy nhin, hin tng nhiu c th xy ra cc gene cch xa nhau ti 30 n v bn ; iu ny ch ra rng cc nhn t khc cng c tm quan trng khng km. Tht l th v, cc tm ng ca cc nhim sc th tm gia rui gim (v c l c nhiu sinh vt khc)

hot ng nh l cc vt cn i vi hin tng nhiu. Ni cch khc, s trao i cho mt vai ca nhim sc th tm gia khng gy hiu qu c ch ln s trao i cho vai kia.
VII. Lp bn bng phn tch b bn (tetrad analysis)

Vic khng nh trao i cho bn si cng thu c Neurospora crassa, mt loi vi nm c nghin cu nhiu trong di truyn hc, c cc sn phm ca gim phn cha ng trong mt dy cc bo t c trt t c sp xp theo kiu thng hng (hnh 4.23). Mt nhn hp t cha trong mt cu trc dng ti gi l nang (ascus), tri qua gim phn hu nh ngay sau khi n va c to thnh. Bn nhn sinh ra t gim phn nm trong mt ci nang theo mt trt t thng hng, v mi nhn ny li tri qua mt ln nguyn phn na to thnh hai bo t nang (ascospores) nm k nhau v ging ht nhau v mt di truyn. Do , mi nang trng thnh c cha tm bo t nang theo bn cp, mi cp biu th mt sn phm ca gim phn. Lc ny, ta c th tch cc bo t nang theo trnh t ca mi nang v cho ny mm trong cc ng nui cy ring xc nh cc kiu gene ca chng.
Bo t v tnh Sinh trng sinh dng
Dung hp Sinh trng sinh dng Hp t Gim phn I Gim phn II Nui cy
Bo t v tnh (bo t dnh)
Ny mm Nguyn phn Nang cha 8 bo t
Ny mm
Hnh 4.23 Vng i ca Neurospora crassa.

Ch thch: Cc bo t sp xp thnh mt dy thng hng trong nang sao cho mi mt na bt ngun t cc th phn ly gim phn I. Mi mt phn t bt ngun t cc chromatid kt hp vi mt tm ng c th.
S sp xp c trt t ca cc sn phm gim phn cho php xc dnh tn s ti t hp gia mt gene v tm ng ca n - y l cch lp bn

tm ng (map the centromere). K thut lp bn ny da trn mt c im ca gim phn, l: cc tm ng tng ng ca cc nhim sc th b m tch ra gim phn I, v cc tm ng ca cc chromatid ch em tch nhau gim phn II. Nh vy, trong trng hp khng c trao i cho gia mt gene v tm ng ca n, cc allele ca gene (v d A v a) tch nhau gim phn I, s phn ly ny c gi l phn ly gim phn I (first-division segregation). Thay v, nu nh mt trao i cho xy ra gia gene v tm ng ca n, cc allele A v a ca n s khng c phn ly cho n lc xy ra gim phn II; s phn ly ny c gi l phn ly gim phn II (second-division segregation). Nh th, khi phn ly gim phn I xy ra, ch c th c hai kiu trt t sp xp ca cc sn phm gim phn, l AAaa v aaAA. Tuy nhin, c th c bn kiu ca phn ly gim phn II, bi v s sp xp ngu nhin ca cc nhim sc th tng ng k gia I v ca cc chromatid k gia II. Bn s sp xp ny c hnh dung nh th ny: AaAa, aAaA, AaaA, v aAAa. T l phn trm ca cc nang c kiu phn ly gim phn II ca mt gene c th c s dng lp bn cho gene gn lin vi tm ng ca n. Chng hn, ta gi s rng c 30% s nang t mt php lai c kiu phn ly gim phn II i vi cc allele A v a. iu ny c ngha l 30% s t bo tri qua gim phn c mt th trao i cho gia A v tm ng ca n. Hn na, trong mi t bo xy ra mt trao i cho, hai trong s cc chromatid l cc th ti t hp v hai ci cn li l nhng th khng ti t hp. Ni cch khc, mt trao i cho n cho ra bn sn phm gim phn, m mt na trong s l cc th ti t hp v mt na l cc th khng ti t hp. Nh vy, tn s trao i cho bng 30% tng ng vi tn s ti t hp l 15%. Theo quy c, khong cch bn phn nh tn s ca cc sn phm gim phn c ti t hp hn l phn nh tn s ca cc t bo m trong trao i cho xy ra. V vy, khong cch bn gia mt gene v tm ng ca n c cho biu thc sau:
(S nang c cc kiu phn ly gim phn II) Tng s cc nang 100
Cng thc ny c gi tr chng no gene nm gn tm ng sao cho mt vi trao i cho phc c th xy ra. Nh th, cc tr s lin kt ng tin cy c xc nh tt nht cho cc gene m chng nm kh gn tm ng. V tr ca cc gene xa hn lc c thc hin bng cch lp bn ca cc gene ny gn lin vi cc gene nm gn tm ng hn. Ti t hp nguyn phn (mitotic recombination) S trao i cho cng c th xy ra trong nguyn phn, mc d vi tn

s thp hn khong 1000 ln so vi gim phn. Bng chng u tin v ti t hp nguyn phn (mitotic recombination) thu c rui gim, nhng li c nghin cu k nht cc vi nm, c bit l nm men bia v Aspergillus. V cc bn di truyn c th thit lp t cc tn s ti t hp trong nguyn phn. Khong cch bn tng i gia cc gene c th i khi tng xng vi khong cch bn da trn cc tn s ti t hp trong gim phn, nhng cha r l do v sao cc khong cch ny li thng khc nhau mt cch ng k. Nhng s tri ngc, khng nht qun ny c th l kt qu hoc l ca cc c ch tip hp khc nhau hoc l ca s phn b khng ngu nhin ca cc v tr trao i tim nng. Khong cch bn v khong cch vt l Nh cp trc y, khong cch bn gia cc gene c c lng bng cc php lai di truyn ni chung l tng ng vi khong cch vt l, hay di ca DNA, m n tch bch cc gene. Mt trng hp ngoi l chnh l vng tm ng, khng c ti t hp nhng c th cha mt di DNA kh ln. Hn na, r rng l ti t hp c th thng c khu tr trong cc vng c bit, v khng phi hon ton ngu nhin trn sut chiu di ca nhim sc th. Bng cch so snh cc bn di truyn (genetic map) v bn vt l (physical map), hnh 4.24 cho thy c khong cch bn c tnh c (bng centiMorgan, vit tt l cM) ln s lng kilobase (1kbp = 1000 bp) trong DNA (bn vt l). K bn bn di truyn ca mi nhim sc th l bn vt l da trn cc nghin cu xc nh bng nhim sc th (hnh 4.24). Bn mi ny c tim nng ht sc to ln trong vic gip cc nh khoa hc xc nh v tr cc gene gy ra cc bnh c th v sau kim tra DNA xem iu g gy ra bnh tt .
Bn di truyn Bn vt l
1. Bn di truyn (2-5 cM) 3. Nhn din gene (to dng cDNA v xc nh trnh t)
Nhim sc th (A)
Trnh t DNA STS 1000 kb). Cc th vin YAC xp gi nhau. (B)
2. Bn vt l (bn
4. Xc nh trnh t (y B gene Ngi vo 4/2003)
Hnh 4.24 Hai cch so snh khong cch bn (A v B); khong cch bn d truyn c cho bng n v cM trn mt nhim sc th v khong cch vt l c cho bng s kilobase trong trnh t DNA.

Gn y, mt n lc to ln c tp trung hng ti thu nhn cho c mt bn di truyn ca b gen ngi (xem hnh 4.24, v cc trang web http://www.genome.gov/ v http://www.ncbi.nlm.nih.gov/Omim/). Bng cch s dng cc ch th phn t (molecular markers) khc nhau (hin gi c bt c ti hng ngn marker tri khp b gene ngi) nh: (i) cc a hnh di phn on DNA (restriction fragment length polymorphism = RFLPs), ngha l cc bin i trong cc trnh t DNA ca cc sinh vt thuc mt loi no m c th xc nh c bng s tch on cc trnh t ny nh s dng cc enzyme gii hn (chng 10), v s bin i nm bn trong v tr gii hn; v RFLPs c th c s dng o a dng ca mt gene trong qun th; hay (ii) V tr trnh t ch (sequence tagged site = STS), l mt trnh t c nht t mt v tr nhim sc th bit m c th khuych i bng phng php PCR; cc STS hot ng nh l nhng marker vt l cho vic lp bn b gene (genome mapping) v to dng (cloning) v.v... Lu : Vn lp bn bng phn tch b bn cc vi nm v vi to tm dng ti y bi v n s c trnh by y trong gio trnh Di truyn hc vi sinh vt v ng dng. Cc hin tng ti t hp bn trong gene (recombination within genes) v php phn tch b tr (complementation) hay l trc nghim cis-trans (cis-trans test) s c cp trong chng 6 v trnh by chi tit trong gio trnh va ni trn.
Cu hi v Bi tp
1. (a) Trnh by cc c ch xc nh gii tnh cc sinh vt, v nu ngha thc tin ca chng. (b) C s di truyn hc ca hin tng bt hot nhim sc th X l g? (c) Phn bit cc tnh trng lin kt vi gii tnh, tnh trng b gii hn bi gii tnh v tnh trng chu nh hng ca gii tnh, v cho cc v d. 2. (a) Ni dung c bn ca thuyt di truyn nhim sc th l g? Tn s tn s ti t hp l g v c ng dng trong di truyn hc nh th no? Cho v d. (b) Hy chng minh: Tn s ti t hp (r) ca hai cp gene bt k trn mt cp nhim sc th tng ng nm trong gii hn 0 r 0,5. C nhn xt g v mi lin h gia cc quy lut di truyn lin kt v phn ly c lp? 3. rui gim, ngi ta tin hnh mt s php lai v nhn c kt qu di y. Hy gii thch c s di truyn cho mi tnh trng v xc nh kiu gene ca cc c th.

P: Ci thn thm, mt trng c thn vng nht, mt F1: Gii ci = tt c vng nht, mt Gii c = tt c vng nht, mt trng F2: 27 vng nht, mt : 25 vng nht, mt trng: 9 thm, mt : 8 thm, mt trng. (Bit rng khng c s khc nhau v t l phn b c ci mi kiu hnh ca F2). 4. Mt php lai gia rui gim ci cnh kha vi rui gim c cnh bnh thng, i con gm c 35 con ci cnh kha, 38 con ci cnh bnh thng v 33 con c bnh thng. (a) Hy xc nh phng thc di truyn ca kiu hnh cnh c kha v s lai. (b) Trn thc t c xut hin rui gim ci cnh kha thun chng hay khng? Ti sao? 5. Khi tin hnh lai gia mt rui gim ci mt v cnh ngn vi rui gim c mt nu v cnh di thu c tt c i con c mt v cnh di. Kt qu tp giao ca cc rui gim F1 thu c F2 nh sau:
Rui gim ci 76 , di 24 , ngn Rui gim c 39 , di 37 nu, di 13 , ngn 11 nu, ngn
Hy xc nh quy lut di truyn ca cc tnh trng v lp cc s lai. 6. Mt ni rui gim kiu di c cho lai vi mt ni khc ng hp v cc th t bin lin kt trn X l m, r v v. Sau em cc rui gim ci F1 lai ngc vi cc con c thuc ni ln th nhn c s lng cc kiu hnh i con ca chng nh di y.
Kiu hnh + m + + + + r + + + + v S lng 331 2 73 14 Kiu hnh m m + m r + +v r v r v S lng 10 81 0 309
Hy xc nh trt t cc gene v lp bn vi khong cch tng i gia cc th t bin ny. 7. Xt cc php lai sau y rui gim v ba gene lin kt vi gii tnh: P: + + +/+ + + y w m/Y F1: Tt c u kiu di

Nu em cc rui gim ci F1 lai phn tch vi cc con c dng kim tra tnh c tn s ca cc kiu hnh nh sau:
Cc lp kiu hnh Nhm b m (khng ti t hp) Nhm ti t hp ca m vi y v w Nhm ti t hp ca y vi w v m Nhm ti t hp ca w vi y v m Tn s 0,664 0,329 0,0057 0,00086
a) Hy xc nh kiu gene ca cc rui gim F1. b) Biu din v tr v khong cch tng i ca cc gene. c) Tnh h s trng hp v nhiu. d) Tnh s c th k vng cho mi kiu hnh i con (bit rng tng s c th l 10.495). 8. Mt rui gim ci d hp v c ba gene +/sc; +/ec; +/vg c cho lai vi mt con c ng hp ln v c ba gene ny. i con gm c:
sc ec vg + + + sc ec + + + vg 233 239 241 231 sc + vg + ec + sc + + + ec vg 12 14 14 16
Hy gii thch kt qu ny v ch ra bn lin kt ph hp. 9. Khi lai hai rui gim Drosophila u d hp v cc tnh trng ln lin kt l mt nu v thn en cho kt qu nh sau: 52 thn vng nht, mt : 9 thn vng nht, mt nu : 10 thn en, mt : 9 thn en, mt nu. Hy gii thch kt qu v lp s lai. 10. rui gim, mt qu thn (k), mt dng mu (cd), v thn mun (e) l ba allele ln lin kt vi nhau. Nu em cc rui ci mt thn, dng mu ng hp lai vi cc rui c thn mun ng hp, thu c cc rui F1 tt c u c kiu di. Nu em cc rui ci F1 d hp lai vi cc con c mt thn, dng mu, thn mun thu c i con gm 1000 con, trong : Mt thn, dng mu 440 Thn mun, mt mu 23 Thn mun 443 Mt thn 25 Mt thn, thn mun 32 Mt thn, mu, mun 1 Mt mu 34 Kiu di 2 a) Xc nh thnh phn gene trn mt nhim sc th ca rui ci F1. b) V bn ca cc gene v tnh h s trng hp v nhiu.

Ti liu Tham kho
Ting Vit Dubinin NP. 1981. Di truyn hc i cng (Bn dch ca Trn nh Min v Phan C Nhn). NXB Nng Nghip, H Ni Hutt FB. 1964. Di truyn hc ng vt. (Bn dch ca Phan C Nhn). NXB Khoa hc v K thut, H Ni, 1978. Phan C Nhn. 2001. Di truyn hc ng vt. NXB Khoa hc v K thut, H Ni. Phan C Nhn (ch bin), Nguyn Minh Cng, ng Hu Lanh. 1999. Di truyn hc (2 tp). NXB Gio Dc, H Ni. Watson JD. 1968. Chui xon kp: Hi k v vic pht minh ra cu trc ca DNA. (Bn dch ca L nh Lng v Thi Don Tnh). NXB Khoa hc v K thut, H Ni, 1984. Ting Anh Ayala FJ, Kiger JA. 1980. Modern Genetics. Benjamin/Cummings Publishing Company, Inc, Menlo Park, CA. Brooker RJ. 1999. Genetics - Analysis and Principles. Benjamin/ Cummings Publishing Company, Inc., Menlo Park, CA. Campbell NA, Reece JB. 2001. Essential Biology. Benjamin/Cummings, an imprint of Addison Wesley Longman, Inc, San Francisco, CA. Clegg CJ, Mackean DG. 2000. Advanced Biology : Principles and Applications. 2nd ed., John Murray Published Ltd, London. Hartl DL, Freifelder D, Snyder LA. 1988. Basic Genetics. Jones and Bartlett Publishers, Inc, Boston - Portola Valley. Kalthoff K. 1996. Analysis of Biological Development. McGraw-Hill, Inc., New York. Karpen GH. 1994. Position-effect variegation and the new biology of heterochromatin. In: Current Opinion in Genetics & Developmment, Vol 4, No 2 (Stillman B and Green M, eds.), pp 281-291. Kimball J. 2004. http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/ Lewis R. 2003. Human Genetics: Concepts and Applications. 5th ed, McGraw-Hill, Inc, NY. Martin E, Ruse M, Holmes E. 1996. Oxford Dictionary of Biology. 3rd. ed., Oxford University Press, Oxford, UK. McKusick V. 1998. Mendelian Inheritance in Man. 12th ed., Johns Hopkins University Press, Baltimore.

Rastan S. 1994. X chromosome inactivation and the Xist gene. In: Current Opinion in Genetics & Developmment, Vol 4, No2 (Stillman B and Green M, eds.), pp 292-297. Russell PJ. 2003. Essential Genetics. Benjamin/Cummings Publishing Company, Inc., Menlo Park, CA. Suzuki DT, Griffiths AJF, Miller JH, Lewontin RC. 1989. An Introduction to Genetic Analysis. 4th ed, W-H Freeman and Company, New York. Tamarin RH. 1999. Principles of Genetics. 6th ed, McGraw-Hill, Inc, NY. Weaver RF, Hedrick PW. 1997. Genetics. 3rd ed, McGraw-Hill Companies, Inc. Wm.C.Browm Publishers, Dubuque, IA. Wellnitz WR. 1995. Genetics: Problem Solving Guide. 2nd ed., Wm.C. Brown Publishers. Dubuque, Iowa. Yablokov AV. 1986. Population Biology (Progress and problems of studies on natural populations). MIR Publishers, Moscow. pp17-30.
Mt s trang web National Human Genome Research Institute (NHGRI)/ National Institute of Health (NIH). 2005: http://www.genome.gov/ http://www.ncbi.nlm.nih.gov/ Online Mendelian Inheritance in Man (OMIMTM): http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=OMIM

Chng 5
Bn cht Ha hc v Ti bn ca Vt cht Di truyn

Trc tin, chng ta cn nm cc c tnh thit yu ca vt cht di truyn sau y: (1) c tnh thng tin sinh hc: cha ng thng tin cn thit cho vic xc nh cu trc ca tt c cc protein c th ca loi (cc gene cu trc) v iu khin cc hot ng sinh trng, phn chia v bit ho t bo; (2) c tnh ti bn: kh nng t sao chp chnh xc, m bo thng tin di truyn ca th h sau ging vi th h trc; (3) c tnh hot ng ca cc gene: cc gene trong b gene c kh nng tng hp ra cc sn phm l nhng phn t tham gia vo mi ng sng cn bn ca t bo (phin m v dch m); (3) c tnh bin i: kh nng b bin i t nhiu qu trnh khc nhau (nh t bin, ti t hp, cc yu t di truyn vn ng). Chnh s bin i a dng ny to ra cc ngun bin d di truyn a dng v phong ph cho s chn lc v tin ho. Ngy nay chng ta u bit rng vt cht di truyn chnh l cc nucleic acid m hu ht sinh vt l loi deoxyribonucleic acid (DNA) v mt s virus l ribonucleic acid (RNA). Trong chng ny, chng ta s tm hiu thnh phn ha hc v cu trc cng nh c ch ti bn ca vt cht di truyn l cc i phn t DNA v RNA.
I. Bng chng vt cht di truyn l cc nucleic acid

1. Cc th nghim bin np vi khun Nm 1928, Frederick Griffith t nn tng cho vic xc nh DNA l vt cht di truyn thng qua th nghim bin np (transformation) trn ph cu khun Streptococcus pneumoniae. Cc vi khun kiu di c v bc bng polysaccharide mc thnh cc khun lc hay dng c trng bng dng nhn (S: smooth); chng c coi l c (virulent) v c kh nng gy vim phi lm chut cht. Mt ni t bin ca n mt kh nng hnh thnh v bc v mc thnh cc khun lc b, x x (R: rough); chng l cc vi khun khng c (avirulent). Griffith tin hnh cc th nghim khc nhau v nhn thy rng: (1) Nu tim cc vi khun sng ni S, chut cht; (2) Nu tim cc vi khun sng ni R, chut sng; (3) Nu tim cc vi khun sng ni S b git cht bng nhit, chut sng bnh thng; (4) Nhng khi tim hn hp gm cc vi khun ni R cn sng v vi khun ni S b git cht bng nhit, kt qu bt ng l chut cht. T mu mu ca cc con chut cht ny ng phn lp c cc vi khun ni S cn sng. iu chng t rng bng

cch no cc mnh v t bo vi khhun ni S b git bi nhit xm nhp vo t bo R sng v lm bin i n thnh vi khun S sng v gy c. Qu trnh ny gi l bin np v cht trong mnh v t bo S gy s bin np c th c gi l tc nhn bin np (transformating agent). Vy bn cht ca tc nhn ny l g?
Ni S sng
Ni R sng
Ni S cht
S cht + R sng
Hnh 5.1 Th nghim ca Griffith (t tri sang l 1-4 nh m t trong bi).
Trong hn mi nm tri k t sau th nghim ca Griffith, Oswald Avery cng vi MacLeod v McCarty tin hnh tr li th nghim bin np ny nhng trong ng nghim (in vitro). H tinh chit cc thnh phn ca tc nhn gy bin np v cho cc enzyme khc nhau tc ng thm d ln hot tnh ca cht chit ny. Kt qu cho thy hot tnh ca cht chit ch b phn hy bi deoxyribonuclease (DNase) - enzyme c kh nng phn ct lm suy thoi ch i vi DNA, m khng b tc ng phn hy ca ribonuclease (RNase) - enzyme lm suy thoi ch cc RNA, cng nh ca cc protease (nh trypsin v chymotrypsin) - enzyme phn gii protein. Vo nm 1944, nhm nghin cu ca Avery chng minh c rng: DNA l vt cht mang thng tin di truyn, ch khng phi protein. 2. Tnh n nh ca hm lng DNA cc sinh vt bc cao Trc khi Avery v cs cng b kt qu ny, a s cc nh ha sinh hc vn tin rng gene chnh l protein. V vy sau kt qu hin nhin , nhiu nh khoa hc vn cn nghi ng tnh ph qut ca n. Vo lc ny, cc nghin cu bn cht ha hc ca nhim sc th khng nh rng, hu ht DNA c khu tr trong cc nhim sc th trong nhn. Hn na, cc phn tch ha hc ca A.E.Mirsky v H.Ris

(USA) v ca R.Vendrely (France) cho thy hm lng DNA mi b nhim sc th lng bi lun lun n nh i vi mt c th nht nh v bng hai ln hm lng DNA mi t bo tinh trng n bi. Chng hn, cc s liu tng ng b l xp x 6,6 v 3,3 picogam (1pg =10-12g). 3. Cc th nghim virus 3.1. Th nghim Hershey-Chase Nm 1952, Alfred D.Hershey v Martha Chase tin hnh cc nghin cu nh du ng v phng x loi th thc khun (bacteriophage, thng ni gn l phage) gi l T2 vn k sinh vi khun Escherichia coli. Phage ny cha mt li DNA v lp v bo v bng protein. Da vo c im nguyn t lu hunh (S: sulfur) ch c trong protein v nguyn t phosphor (P) ch c trong nucleic acid, h nh du protein bng cht ng v phng x S35 v DNA bng P32 bng cch cho cc phage ny sinh trng trn cc mi trng c S35 v sinh sn khi c S32. Cc virus nh du c s dng theo di thnh phn no t virus ban u i vo t bo ch v sau xut hin trong cc phage th h con.
Ht virus
DNA V virus
Hnh 5.2 Th nghim Hershey-Chase. (a) Phage bm b mt ngoi mng t bo vi khun; (b) bm li DNA vo trong v li lp v protein bm ngoi; (c) ti bn, phin m v tng hp hng lot protein ca virus; (d) lp rp li + v to ra cc virion; (e) enzym lysozyme ph hy vch t bo vi khun v phng thch cc phage th h con.
Kt qu cho thy trong cc phage th h con ch c hu nh DNA dng ban u v khng h c protein dng ny (hnh 5.2). Hn na, lp v protein dng ban u bm bn ngoi mng t bo vi khun, khng thc hin chc nng no sau khi DNA c tim vo trong vi khun. Nh vy, vt cht di truyn ca phage T2 l DNA, ch khng phi protein. 3.2. Th nghim chng minh vt cht di truyn ca mt s virus l RNA

Sau ny, nm 1956 A.Gierer chng minh rng RNA c tinh sch t virus m thuc l (tobacco mosaic virus = TMV) c kh nng ly nhim khi khng c bt k protein no thuc virus. Kt qu phn tch cc virus th h con cho thy chng hon ton ging vi cc virus sinh ra t s cho ly nhim bng cc virus cn nguyn vn. Bng chng RNA l thnh phn di truyn ca TMV cng c Fraenkel Conrat v B.Singer ti xc nhn nm 1957 trong th nghim "lp rp cho" gia li RNA v v protein ca hao kiu TMV l S v HR.
II. Thnh phn ha hc v cu trc ca DNA

Cc nucleic acid (DNA, RNA) l nhng polymer sinh hc, c trng lng phn t ln, gm nhiu n phn (monomer) l cc nucleotide ni vi nhau to thnh cc chui hay mch polynucleotide - cu trc s cp ca cc phn t nucleic acid. 1. Cu trc ca cc nucleotide
lin kt glycosid
Hnh 5.3 Bn loi base ca DNA v cu trc mt nucleotide (dAMP).
Mi nucleotide gm ba thnh phn: mt nhm phosphate, mt gc ng pentose v mt trong bn loi base nit (cc dn xut ca purine hoc pyrimidine). V cu trc, nhm phosphate ni vi gc ng ti nguyn t carbon s 5 (C5') bng mt lin kt ester v base nit ni vi gc ng ti nguyn t carbon s 1 (C1') bng mt lin kt -glycosid (Hnh 5.3). Lu : (1) Cc base l thnh phn c trng qui nh tn gi cc nucleotide mang chng. Trong DNA cha bn loi base c bn:

adenine (A), thymine (T), guanine (G) v cytosine (C); trong RNA cng cha bn loi nhng ch khc l thymine c thay bi uracil (U). (2) ng pentose ca RNA l D-ribose v ca DNA l 2-deoxy-Dribose (k hiu D-: ch dng ng quay phi phn bit vi dng Lquay tri khng c trong thnh phn ca cc nucleic acid t nhin). Hai gc ng ny khc nhau C2'; trong ribose l nhm -OH v trong deoxyribose l -H. S c mt ca thnh phn ng trong cc nucleotide phn bit hai loi ribonucleotide v deoxyribonucleotide cu to nn hai loi nucleic acid tng ng l RNA v DNA. (3) Cu trc "base + ng" gi l nucleoside; cch c v vit tt tng ng vi cc base A, T, G v C trong DNA nh sau: deoxyadenosine (dA), deoxythymidine (dT), deoxyguanosine (dG) v deoxycytidine (dC); cch gi cho cc nucleoside trong RNA ch cn b tin ng "deoxy" (d), v thay cho dT l uridine (U). V nu c y tn gi ca mt nucleotide ch cn thm ci "ui" 5'-monophosphate, v d hnh 5.3. (4) Tnh phn cc trong cu trc mt nucleotide cn th hin cc nhm hydroxyl (-OH) hai v tr C5' (hnh thnh lin kt ester vi nhm phosphate to ra nucleotide) v C3' (hnh thnh lin kt phosphodiester vi nucleotide khc to chui polynucleotide). 2. Cu trc chui polynucleotide
u 5'
Lin kt 3',5'-phosphodiester v chiu 5'3' ca chui polynucleotide
u 3'
Hnh 5.4 Cu trc chui polynucleotide ca DNA.

Cc nucleotide trong DNA hoc RNA ni vi nhau bng cc mi lin kt 3',5'-phosphodiester gia gc ng ca nucleotide ny vi nhm phosphate ca nucleotide k tip, to thnh chui polynucleotide (nh s xc tc ca cc enzyme tng ng l DNA- hoc RNA-polymerase). V vy cc chui ny bao gi cng c ko di theo chiu 5'3' (u 5' mang nhm phosphate t do v u 3' cha nhm -OH t do) v c b khung gm cc gc ng v phosphate lun phin nhau, cn cc base nh vy c trnh t c c theo mt chiu xc nh (hnh 5.4). Thng thng ngi ta biu din trnh t base 5'3' theo chiu t tri sang phi. V d di y cho thy cc chui RNA v DNA ch khc nhau bi base U v T v gc ng trong cc nucleotide ca chng. chui RNA 5'- AUAGACUACG-3' chui DNA 5'- dAdTdAdGdAdCdTdAdCdG-3' 3. Thnh phn ha hc v cu trc ca chui xon kp DNA 3.1. Thnh phn ha hc ca DNA Nm 1949, E.Chargaff p dng phng php sc k giy vo phn tch thnh phn ha hc ca DNA cc loi khc nhau (Bng 5.1; c th xem trong Watson et al 1987, tr.73) khm ph ra ba im quan trng sau y: (1) S lng bn loi base trong DNA l khng bng nhau; (2) T l tng i ca cc base l khng ngu nhin; v trong tt c cc mu DNA nghin cu tn ti mi tng quan v hm lng (%) gia cc base nh sau: A T v G C, ngha l t s (A+G)/ T+C) 1; v (3) Mi loi c mt t l (A+T)/(G+C) c th.
Bng 5.1 Thnh phn base ca DNA mt s loi (t nhiu tc gi) Sinh vt Phage lambda Phage T7 Mycobacterium tuberculosis Escherichia coli Aspergillus niger (nm mc) Saccharomyces cerevisiae Triticum (la m) Zea mays (ng) Salmo salar (c hi) Gallus domestica (g nh) Homo sapiens (ngi) A% 21,3 26,0 15,1 24,7 25,0 31,3 27,3 26,8 29,7 29,5 30,9 T% 22,9 26,0 14,6 23,6 24,9 32,9 27,1 27,2 29,1 27,7 29,4 G% C% 28,6 24,0 34,9 26,0 25,1 18,7 22,7 22,8 20,8 22,4 19,9 27,2 24,0 35,4 25,7 25,0 17,1 22,8 23,2 20,4 20,4 19,8
A+G T +C
1,00 1,00 1,00 1,03 1,00 1,00 1,00 0,98 1,02 1,08 1,01
A+T G+C
0,79 1,08 0,42 0,93 1,00 1,79 1,19 1,17 1,43 1,34 1,52

3.2. Cu trc chui xon kp Vo nm 1951-52, vic nghin cu cu trc ba chiu ca DNA bng phn tch nhiu x tia X c bt u bi Maurice Wilkins v Rosalind Franklin. Cc bc nh chp c 1952 (hnh 5.5) gi rng DNA c cu trc xon gm hai hoc ba chui. Lc ny Anh cn c mt s nghin cu khc nhm pht trin l thuyt nhiu x ca Linus Pauling tm hiu cu
Hnh 5.5 nh chp DNA tinh th bng tia X ca Franklin.
trc DNA. Tuy nhin, gii php ng n nht l chui xon kp (double helix) b sung do James Watson v Francis Crick a ra nm 1953. M hnh ny (hnh 5.6) ph hp vi cc s liu ca Wilkins v Franklin cng nh ca Chargaff. S kin ny m ra mt bc ngot mi cho cho s ra i v pht trin vi tc nh v bo ca di truyn hc phn t v sinh hc ni chung.
Lin kt hydro B khung ng-phosphate
Hnh 5.6 Cc m hnh cu trc DNA. Bn tri l m hnh khng gian lp y. Hnh gia l chui xon dui thng cho thy cc cp base gia v hai b khung ng-phosphate hai bn, vi cc thnh phn c ch ra bng cc mi tn. Bn phi l s chui xon kp vi hai si i song song.
M hnh Watson-Crick hay DNA dng B c cc c im sau:

(1) DNA gm hai chui i song song (antiparallel) cng un quanh mt trc trung tm theo chiu xon phi, vi ng knh 20Ao (1angstrom = 10-10m), gm nhiu vng xon lp li mt cch u n v chiu cao mi vng xon l 34 Ao, ng vi 10 cp base (base pair, vit tt l bp). (2) Cc b khung ng-phosphate phn b mt ngoi chui xon v cc base nm bn trong; chng xp trn nhng mt phng song song vi nhau v thng gc vi trc phn t, vi khong cch trung bnh 3,4 Ao. (3) Hai si n gn b vi nhau bng cc mi lin kt hydro (vn l lc ha hc yu) c hnh thnh gia cc cp base i din theo nguyn tc b sung "mt purine - mt pyrimidine". C th l, trong DNA ch tn ti hai kiu kt cp base c th l A-T (vi hai lin kt hydro) v G-C (vi ba lin kt hydro) (xem cc hnh 5.6 v 5.7). (4) Tnh cht b sung theo cp base dn n s b sung v trnh t cc base gia hai si n ca mi chui xon kp. V vy, trong DNA si kp bao gi cng c: A = T v G = C (quy lut Chargaff); ngha l (A+G) = A+T A+G c th cho tng loi. (T+C) hay t s = 1 , cn t l T +C G+C
Hnh 5.7 Hai kiu kt cp base ca DNA. Cp G-C ni vi nhau bng ba lin kt hydro (biu th bng cc ng chm), c cng hnh dng chnh xc nh cp A-T ni vi nhau bng hai lin kt hydro. Cc mi tn ch v tr lin kt gia base vi gc ng.
Cn lu rng, hai c im quan trng nht trong cu trc DNA l s phn cc ngc chiu ca hai si n (5'3' v 3'5') v nguyn tc b sung gia cc cp base. Hai c im ny cho php gii thch mt cch c

bn cc c ch di truyn cp phn t (ti bn, phin m v dch m). 4. S lc v cc c tnh ha l ca cc nucleic acid 4.1. Cc dng ca DNA M hnh Watson-Crick hay DNA dng B l cu trc ph bin. Tuy nhin, sau ny ngi ta cn pht hin ra nhiu dng xon phi khc (A,C,D...); chng c mt s bin i so vi DNA-B (xem bng 5.2). Bn cnh cc dng DNA xon phi, t nm 1979 Alexander Rich v ng s cn pht hin thm mt dng DNA xon tri duy nht cho n nay, gi l DNA-Z. Dng DNA ny c cc b khung zigzag un gp khc theo chiu tri, mi vng xon di 45,6Ao cha 12 cp base (hnh 5.8 v bng 5.2). DNA dng Z cha cc si gm cc purine v pyrimidine xp xen k nhau, th d: --GCGCGCGC---CGCGCGCG-Mc d Rich khm ph DNA-Z khi nghin cu v cc hp cht m hnh, song cu trc ny dng nh cng c mt trong cc t bo sng mt t l nh v vn cn cha tht s hiu r chc nng ca n. Hnh 5.8 DNA-B v DNA-Z
Bng 5.2 c im ca cc dng DNA - A, B, C v Z Dng Chiu xon S bp/vng xon ng knh chui xon A Phi 11,0 23Ao B Phi 10,0 19Ao C Phi 9,3 19Ao Z Tri 12,0 18Ao *Ngun: J.Kimball (t internet). V chi tit, xem trong Watson et al 1987, tr.249.
4.2. Bin tnh v hi tnh ca DNA 4.2.1. Khi nim bin tnh v hi tnh Mt trong nhng c im quan trng nht ca DNA l hai mch n b sung ca n gn vi nhau bng cc mi lin kt hydro, vn l lc lin kt ha hc yu nn chng c th b phn hy di tc dng ca cc enzyme, nng lng... lm cho hai mch n ca chui xon kp tch ri nhau. Nh DNA mi c th ti bn, cc gene mi c th biu hin ra

cc sn phm ca mnh. Mt khc, DNA c th phc hi tr li trng thi ban u theo mt qu trnh ngc li, ngha l c tnh thun-nghch. Bng thc nghim, ngi ta chng minh iu bng cch s dng cc tc nhn vt l v ha hc khc nhau. Chng hn, khi un nng t t cc phn t DNA ln ti nhit gn 100oC (thng l 90-95oC), th cc lin kt hydro ca chng b ph hy hon ton v hai si b sung tch ra. Hin tng ny c gi l bin tnh (denaturation). Ngc li, khi lm ngui t t dung dch t nng cha DNA b bin tnh han ton, cc si n thng cp i vi si b sung ca chng v lm phc hi chui xon kp nh lc u. Hin tng gi l hi tnh (renaturation).
DNA si kp giu GC ch- a bnng chy
Tm ph thuc vo hm l- ng G-C ca DNA

% hyperchromicity
50
V ng DNA giu A-T - c t ch thnh mt bp d ng s i n
DNA E. coli v i 50% G-C, c Tm bng 69 o C.
60
70
80
C c v ng giu A-T t ch tr- c, c c v ng giu G-C t ch sau
Nhi oC t Hm l- ng G-C c th- c x c t Tm ca DNA nh
Hnh 5.9 DNA bin tnh tng phn. Hnh 5.10 S ph thuc ca Tm vo hm lng GC ca 3 DNA khc nhau
nhit va phi hoc khi c mt cc tc nhn gy mt n nh nh alkali hay formamide, th cc phn t DNA b bin tnh tng phn. Khi ti cc vng giu cp AT s tch tng phn trc, trong khi cc vng giu cp GC vn gi nguyn c tnh xon kp. iu ny c th l gii l do mi cp AT ch c hai lin kt hydro r rng l km bn hn so vi mi cp GC c ti ba mi lin kt nh th. Nhit m ti cc si DNA b bin tnh hay tch nhau mt na c gi l nhit nng chy (melting temperature), hay Tm. Tm l im gia ca pha phuyn tip (hnh 5.9) v n ty thuc vo hm lng G-C ca DNA, ngha l c trng cho DNA mi loi. V d, DNA ca E. coli vi 50% G-C th c Tm l 69oC (hnh 5.10). Ni chung, hm lng GC ca mt DNA c th bin thin t 22% nm mc nhy Dictyostelium n 73% Mycobacterium phlei. iu ny c th gy mt hiu qu mnh ln cc c tnh ha l ca DNA, c bit l ln nhit nng chy, vn n tng tuyn tnh vi hm lng GC. Nhit nng chy (Tm) ca mt DNA l nhit m ti hai si b bin tnh hay tch nhau mt na. Ngoi ra, nng ion thp v cc dung mi hu c cng thc y s bin tnh ca DNA.

4.2.2. ng dng trong lai phn t (hnh 5.11) Ngi ta li dng kh nng ni trn ca DNA to ra cc phn t DNA lai nhn to bng cch lm lnh t t hn hp cc DNA bin tnh t hai loi khc nhau. K thut lai phn t (molecular hybridization) ny c ng dng rng ri xc nh mc tng ng DNA ca cc nhm phn loi khc nhau. V d, cc thc nghim cho thy c khong 25% tng s DNA ngi v chut c th lai vi nhau (Watson et al 1987).
Hnh 5.11 Bin tnh v hi tnh ca DNA (tri) v lai nucleic acid (phi).
Theo ngha rng, lai phn t hay lai nucleic acid c hiu l s tng tc gia cc si nucleic acid b sung. N c th xy ra gia hai si DNA hay gia cc si DNA v RNA, v chnh l c s ca nhiu k thut, chng hn nh: lai mt mu d c nh du ng v phng x (radioactive probe) lc ra DNA hay RNA bm vo mang lai l mt trong nhng th nghim cho nhiu thng tin b ch nht c tin hnh trong di truyn hc phn t. Hai kiu c bn ca lai phn t l phng php thm Southern (Southern blots) v phng php thm Northern (Northern blots). Trong , kiu u l lai bng mt mu d lc DNA, cn kiu sau dng lc RNA. Mu d (probe) l cc cng c s cp c dng xc nh cc trnh t b sung cn quan tm; n l mt nucleic acid si n c nh du phng x v c dng xc nh mt trnh t nucleic acid b sung bm trn mng lc nitrocellulose hay mng lai bng nylon. Ni chung, mu d l mt dng (clone) c to ra bng cch xen DNA vo mt vector. Thng thng hu ht cc vt d ny l cc dng thuc plasmid.
III. Kch thc b gene v tnh phc tp v mt tin ha

1. S lc v b gene ca cc virus, prokaryote v eukaryote Virus l nhm "sinh vt" b nht cha c cu to t bo, khng tn ti n c m k sinh bt buc cc t bo sinh vt tin nhn (prokaryote) hoc sinh vt nhn chun (eukaryote); ch trong iu kin chng mi c kh nng ti bn. Cc virus k sinh hoc gy nhim vi khun gi l th

thc khun (bacteriophage) hay phage. Cu trc ca cc virus tng i n gin, gm hai phn chnh l li axit nucleic v v protein (hnh 5.12). Mt s virus thc vt hoc cc virus gy ung th, AIDS, SARS ngi v ng vt c b gene l RNA. S cn li bao gm nhiu virus k sinh vi khun v ng vt c b gene l DNA si kp hoc si n, c mch thng hoc mch vng (bng 5.3).
Cc enzyme RNA V (capsid) Bao virus
Protein virus
Hnh 5.12 nh chp phage T4 (tri) v s cu trc ca HIV (phi).
Hnh 5.13 nh chp cc t bo E. coli (tri) v vt cht di truyn ca n (phi), v bn di l nh hin vi in t ca plasmid pSC101.
Nhm prokaryote bao gm cc vi khun (bacteria) v vi khun c (archae) l cc sinh vt c cu to t bo n gin nht. Vi khun Escherichia coli (hnh 5.13) l i tng c s dng rng ri trong cc nghin cu di truyn phn t. B gene chnh ca n l mt phn t DNA si kp vng c kch thc ln (4.639.221 bp, vi 4290 gene m ha protein v 53 gene RNA) gi l nhim sc th chnh. N thng tp trung mt "vng nhn" (nucleoid), khng c mng nhn bao bc, v trng thi siu xon (supercoiled DNA) di s kim sot ca cc enzyme

topoisomerase. Ngoi ra cn c nhiu phn t DNA si kp trn mch vng khc c knh thc b hn nhiu, gi l cc plasmid (Vn ny c trnh by k trong gio trnh Di truyn hc Vi sinh vt v ng dng). Nhm eukaryote l nhm ln nht v tin ha a dng nht v trnh d t chc c th, bao gm tt c cc sinh vt c cu to t bo (tr vi khun v vi khun lam), c th l n bo hoc a bo. Trong t bo cha hai h thng di truyn, b gene nhn ( trnh by chng 3 v 4) v b gene t bo cht (xem chng 9). Kch thc b gene ca mt s sinh vt thng gp c gii thiu cc bng 5.3 v 5.4. 2. Mi quan h gia kch thc b gene v tnh phc tp v tin ha Da trn cc kt qu phn tch b gene ca cc virus v vi khun cng nh ca b gene eukaryote (b nhim sc th n bi), cho php khi qut nh sau: Kch thc ca b gene tng ln tng i cng vi mc phc tp v tin ha. Tht vy, t bng 5.3 ta thy rng kch thc b gene ca cc virus ni chung l rt nh so vi nhm prokaryote; v kch thc b gene ca cc prokaryote li t ra qu n gin so vi ngay c mt eukaryote n bo nh nm men. V d: Trong khi DNA ca mt vi khun in hnh nh E. coli hn 4,6 triu cp base v cha 4.377 gene m ha protein (khng k 53 gene RNA); th phage MS2 l mt trong cc virus b nht, b gene RNA ca n cng ch c 3.569 base vi tt c 4 gene; hoc c kch thc ln nh virus Epstein-Barr - b gene DNA si kp mch vng - cng ch c 172.282 cp base vi tt c 80 gene. Nu xt trn c hai nhm prokaryote v eukaryote, ta thy rng kch thc cc b gene bin thin rt rng: b gene i vi mt sinh vt sng t do (mt vi khun) c bit l b nht cha khong 600.000 cp base DNA, trong khi cc b gen ngi v chut l khong 3 t. Nu xt ring nhm eukaryote, ta bit mi loi c mt s lng nhim sc th c trng v n khng phn nh trnh tin ha ca cc loi. Tuy nhin, v mt no r rng l c s tng quan thun gia hm lng DNA ca cc b gene n bi v nc thang tin ha ca cc ngthc vt. D vy vn c mt s ngoi l so vi quy tc ny! 3. Hm lng DNA v nghch l gi tr C Chng hn, mc d Psilotum nudum, i khi gi l "dng x lng", l mt thc vt n gin hn nhiu so vi loi Arabidopsis thaliana vn l mt thc vt c hoa thuc h ci, nhng n li c hm lng DNA ln hn ti 3.000 ln. Mc d ta hiu ti sao, nhng c iu th v l trn 80% b gene ca n l DNA lp li khng cha thng tin di truyn no c! Hay mt s thc vt (nh ng, loa kn) hay mt s ng vt (nh lng th, c) li c kch thc b gene ln gp nhiu ln so vi lp th (bng 5.3).

Mt s lng th cha DNA nhiu gp b gene chng ta n 30 ln, nhng chc chn khng phi l chng phc tp gp chng ta 30 ln.
Bng 5.3 Kch thc b gene (genome) mt s sinh vt thng gp B gene sinh vt Virus Phage -X174 ( E. coli) Phage lambda ( E. coli) Phage T2 hoc T4 ( E. coli) Phage MS2 ( E. coli) SV40 (gy khi u kh) Virus SARS (v d, H5N1) Epstein-Barr virus (EBV) Prokaryote Haemophilus influenzae Chlamydia trachomatis Streptococcus pneumoniae Vibrio cholerae Mycobacterium tuberculosis Bacillus subtilis Escherichia coli* Agrobacterium tumefaciens** E. coli O157:H7 Eukaryote Saccharomyces cerevisiae Schizosaccharomyces pombe Plasmodium falciparum*** Neurospora crassa Caenorhabditis elegans**** Arabidopsis thaliana Drosophila melanogaster Ngi (Homo sapiens)***** La (Oryza sativa ) Loa kn (Lilium longiflorum) Chut Lng th S bp 5.386 48.502 ~2 105 3.569 5.226 29.600 172.282 1.830.138 1.042.519 2.160.837 4.033.460 4.411.532 4.214.814 4.639.221 4.674.062 5,44 106
S gene
Ghi ch
10 DNA si n vng ~61 DNA si kp thng 150-200 DNA si kp thng 4 RNA DNA si kp vng RNA 80 DNA si kp thng 1.738 936 2.236 3.890 3.959 4.779 4.377 5.419 5.416 Gy nhim tai gia Bnh ly qua tnh dc Pneumococcus
2 NST; gy cholera
Gy bnh lao C 4290 cistron Vector hu ch... Gy bnh ngi Men bia ny chi Nm men phn ct Gy st rt nguy hi + 498 gene RNA Gii tr.t u tin... Thc vt c hoa Rui gim Gii tr.t 4/2003 ...
5.770 12.495.682 12.462.637 4.929 22.853.764 5.268 38.639.769 10.082 100.258.171 ~19.000 25.498 115.409.949 122.653.977 13.379 ~3,2 109 ~25.000 4,3 108 ~60.000 3 1011 ? 2,2 109 ? ? 109 - 1011
Ch thch: *C 4290 gene m ha protein, cn li l cc gene RNA; ** Vector hu ch chuyn gene thc vt; *** Cng vi 53 gene RNA; **** Eukaryote a bo u tin c xc nh trnh t; ***** c xc nh y trnh t vo 4/2003 bi HGP, vi tng cng 3.164.700.000 cp base; v theo c tnh mi nht cng b ngy 21/10/2004 trn tp ch Nature, b gene ngi chng ta cha khong 20.000-25.000 gene m ha protein, chim khong 2% ton b b gene.

Ngi ta gi tng hm lng DNA trong b gene n bi l gi tr C (C-value). Vi phn tch trn cho thy khng h tn ti mt mi quan h nht qun gia gi tr C v tnh phc tp ca mt sinh vt (nh lng th vi th); ci c gi l nghch l gi tr C (C-value paradox). 4. V kch thc DNA cc bo quan Kch thc DNA ca mt s bo quan (bng 5.4) cho thy chng c v n gin v khng c du hiu tin ha r rt. Ni chung, kch thc mi phn t DNA ty th (mitochondrial DNA = mtDNA) ca ngi v cc ng vt c v thng nm trong khong 15.000-17.000 bp; v d mtDNA ngi l 16.569 bp. Trong khi , kch thc mt DNA lp th (chloroplast DNA = ctDNA hay cpDNA) phn ln t bo cc thc vt thng vo khang 130.000 -150.000 bp. Chng hn, ctDNA la trng (O. sativa) thuc hai nhm indica v japonica c kch thc tng ng l 134.494 bp v 135.525 bp, la m (Triticum aestivum) l 134.545 bp v ng (Zea mays) l 140.384 bp...Cn cc plasmid hin din mt s t bo thc vt thng c kch thc rt b khong 1-2 ngn cp base.
Bng 5.4 Kch thc DNA bo quan mt s sinh vt DNA ty th Ngi D. melanogaster S. cerevisiae Plasmid La (indica; japonica) Ng S cp base 16.569 19.517 85.779 1485 ; 2.135 1.913 DNA lp th La (ind; jap) Ng Ma Plasmid N. crassa Brassica S cp base 134494 ; 134525 140.384 141.182 3581; 3675;7050 11.640
IV. T chc phn t ca cc nhim sc th

1. Cu trc cht nhim sc y ta ch tm hiu t chc ca DNA trong cc nhim sc th eukaryote. Mi nhim sc th eukaryote l mt phc hp nucleoprotein bao gm mt phn t DNA si kp mch thng kt hp vi cc phn t protein c s gi l histone (giu cc amino acid lysine v arginine). Phc hp nh th gi l cht nhim sc (chromatin).
Hnh 5.14 S cu trc mt nucleosome

n v t chc ca c s cc nhim sc th eukaryote l cc nucleosome (hnh 5.14). Mi nucleosome c ng knh khong 11 nm, gm mt khi cu tm phn t histone, (H2A+ H2B +H3+H4)2, gi l li octamer v on DNA c kch thc 146 cp base qun 1 vng xung quanh n (thng c m t n gin: mt on DNA di 160-200 bp qun hai vng quanh octamer). Mt phn t H1 bm vo on DNA "ni" (linker DNA) bn ngoi khi cu, gi vng s tng tc ca DNA vi cc histone li. on DNA ni gia hai nucleosome di khong150 bp.
DNA cun cht
trong NST k gia
on DNA si kp Chui nucleosome
NST k gia
Chromatin c c
Si chromatin
30 nm
Cc nucleosome
Vng cun vng
Chui xon kp DNA
Vng xon cht
NST k gia
Hnh 5.15 T chc DNA trong nhim sc th eukaryote. DNA cun cht trong nhim sc th k gia c m xon dn qua cc mc khc nhau (hnh tri). DNA ng xon dn qua cc mc cho n khi hnh thnh nhim sc th k gia nguyn phn, vi chiu di rt ngn khong 50.000 ln (hnh phi).
Nh vy, bc cu trc u tin ca chromatin c hnh dung di dng mt chui cc nucleosome, hay si nucleosome (nucleosome fiber) c dy 11 nm (1 nanomet = 10 Ao). Bc th hai ca ca s cun gp chromatin c lin quan ti s xon li ca si nucleosome thnh ra mt

si dy 25 nm gi l mt solenoid. Cc histone H1 c coi l tham gia vo s xon li ny bng cch tng tc vi cc phn t H1 khc. Bc th ba ca s ha xon c l l cun vng ca si 25 nm thnh mt cu trc kiu nh bn chi (brushlike) vi cc vng c neo dnh vo mt gi trung tm (dy ~300 nm). y chnh l vng tho gin xon ca nhim sc th, tng ng vi cht ng nhim sc (euchromatin). Sau cc dy vng c sp xp trong cc khng gian ba chiu ny xon cht to thnh cc vng gi l cht d nhim sc (heterochromatin) trn mt chromatid vi dy khong 700 nm. Ti k gia ca nguyn phn, mi nhim sc th c cu trc in hnh gm hai chromatid ch em (two sister chromatids) dnh chung nhau tm ng (centromere) vi dy ton b chng 1400 nm (hnh 5.15). Nh vy, cht nhim sc trong cc t bo eukaryote tn ti hai dng: (1) Cht d nhim sc l cc phn cun xon cht v khng c hot tnh phin m; v (2) Cht ng nhim sc l cc vng gin xon v ch t cng c tim nng hot tnh. 2. S lc cc thnh phn DNA trong b gene eukaryote Ni chung, trong mi b gene eukaryote bao gm cc kiu DNA chnh sau y, v cho tin ta ly b gene ngi lm th d : - DNA bn sao n (single-copy or unique), ngha l cc gene c mt ch mt ln trong b gene; loi ny chim khong 75% b gene v bao gm hu ht cc gene. - DNA lp li (repetitive) bao gm mt s gene c lp li vi ln cho n hng ngn ln trong b gene. Nhm ny c chia lm hai loi: + DNA lp li kiu phn tn (interspersed): loi ny chim khong 15% v phn b ri rc khp b gen gia cc gene cng nh bn trong cc gene; bao gm cc trnh t Alu (l cc on DNA di khong 300 bp v c lp li khong 300.000 ln trong b gene; chng c th c mt ch tip gip hoc bn trong cc gene trong cc intron hoc cc vng khng c dch m) v c cc on lp ni tip c s lng bin thn (VNTRs = variable numbers of tandem repeats), vn l cc on lp ngn ch vi cp base nhng c chiu di sai khc, hay cc tiu v tinh (mini- hay microsatellite). V d: cc on lp phn tn (5'3') trong cc on Alu: GCTGCGG........GCTGAGG........GCTGAGG + DNA v tinh (satallite) hay DNA lp li kiu ni tip (tandem): loi ny chim khong 10% v c lp li rt nhiu ln, bao gm cc trnh t n gin thng khu tr cc tm ng (centromere) v cc u mt (telomere) ca cc nhim sc th. V d: cc on lp ni tip (5'3') thuc cc microsatellite hay telomere:

5'-...TTAGGGTTAGGGTTAGGGTTAGGG...-3' Nhn chung, b gene ngi c cc c im sau: (1) V b gene nhn, kch thc b gene n bi l ~3,2 t bp; hu nh tt c mc phc tp ca n u nm trong lp DNA bn sao n (~75%). B gene ngi c coi l c cha khong 25.000 gene m ha protein. Trong a phn l cc gene phn on vi cu trc phc tp (xem chng 6). Cc gene u cha t 1 cho n trn 75 exon (on m ha protein) v c chiu di bin thin t di 100 cho n khong 2.400.000 bp (gene gy ri lon dng c DMD c xem l di nht vi 2,4 triu bp ny chim ~ 1,5% nhim sc th X m n nh khu). Trnh t Alu c mt khp b gene. (2) V b gene ty th, y l b gene mch vng vi 16.569 bp v cha ~ 40 gene.
V. Ti bn DNA
Sau khi khm ph ra cu trc DNA, Watson v Crick xut ba kiu truyn thng tin di truyn c th c trong cc t bo: (1) Ti bn (replication): DNADNA; (2) Phin m (transcription): DNARNA; v (3) Dch m (translation): RNAProtein. Cc knh truyn thng tin di truyn ny c gi l Gio l Trung tm (Central Dogma) ca sinh hc phn t. Sau ny n nm 1970, Baltimore v Temin qua nghin cu c ch hot ng ca b gene RNA virus Sarcoma b sung thm knh RNADNA, gi l phin m ngc (reverse transciption) v n ngc hng vi knh phin m ph bin. iu ny c minh ha hnh 5.16.
Dch m
Phin m Phin m ngc
Ti bn
Hnh 5.16 Cc kiu truyn thng tin di truyn (tri), v m hnh ti bn DNA theo kiu bn bo ton do Watson v Crick xut.
1. Cc nguyn tc v c im chung ca ti bn DNA Ti bn (replication) l mt c tnh quan trng nht ca vt cht di truyn, nh s sng c duy tr lin tc, cc loi bo tn c tnh cht c trng ca mnh, v con ci thng ging b m. Vy DNA v cc b gene ni chung c ti bn nh th no?

Trong khi khm ph ra m hnh cu trc DNA, chnh Watson v Crick a ra d on chnh xc (da trn nguyn tc b sung ca cc cp base) rng s ti bn DNA phi din ra theo kiu bn bo ton (semiconservative) nh hnh 5.16. n 1956, S. Luria v Max Delbruck ngh ba kiu ti bn c th c: bn bo ton, bo ton (conservative) v phn tn (dispersive). Tuy nhin, nhiu th nghim sau nhanh chng khng nh s ti bn DNA din ra theo kiu bn bo ton. in hnh l th nghim ca Meselson v Stahl nm 1958 trn i tng l E. coli bng phng php nh du ng v phng x N15 (nit nng) kt hp vi ly tm siu tc (ultra-centrifugation). u tin cho vi khun ny sinh trng trn mi trng N15; ri a tr li mi trng N14 (nit nh) v sau mt, hai hoc ba th h u c ly cc mu DNA. tch DNA nng v nh, ngi ta cho trn ln cc mu ny vi cesium chloride (CsCl) trc khi em ly tm. Khi ly tm, DNA c cc t trng nng (heavy), nh (light) v trung bnh (intermediate) s tch thnh cc vch tng ng khc nhau trong ng nghim (hnh 5.17).
DNA cha m
Hnh 5.17 Th nghim Meselson-Stahl v s ti bn bn bo ton ca DNA.
Cc kt qu cho thy rng sau mt th h, 100% DNA si kp c t trng trung bnh, ngha l mt si nng (t dng cha m) v mt si nh (c tng hp mi). Kt qu ny cho php d an v kt lun s ti bn xy ra theo kiu bn bo ton ng nh d on ca Watson v Crick rng, cc si DNA lm khun cho s ti bn ca ring chng. Di y l cc nguyn tc v c im chung ca ti bn DNA. (i) Ti bn theo kiu bn bo ton v gin on (discontinuous); (ii) S ti bn c bt u ti mt hoc nhiu v tr c th trn phn t DNA v din ra ng thi theo hai hng ngc nhau gi l khi im ti bn hai hng (bidirectional origin of replication). Ngha l, t khi im ny DNA si kp m xon thnh hnh vng m sinh trng theo hai hng i lp nhau to ra hai chc ti bn (replication fork). Mi khi im ti bn cng vi hai chc nh vy goi l mt n v ti bn

(replicating unit hay replicon) c minh ho hnh 5.18. Ni chung, i vi cc DNA mch vng (b gene mt s virus, vi khun, cc plasmid v cc DNA bo quan ca eukaryote), mi phn t ch c mt khi im ti bn; trong khi mi DNA trong nhim sc th eukaryote c nhiu khi im ti bn hot ng theo mt trnh t c th (xem hnh 5.20).
khi im (Ori)
chc
chc
sinh trng
sinh trng
Hnh 5.18 Mt khi im v hai chc ti bn sinh trng theo hai hng i lp nhau. y cng cho thy on mi RNA c tng hp trc.
(iii) Qu trnh ti bn DNA ph thuc vo mt h thng gm nhiu protein v enzyme khc nhau (xem mc 2 bn di); (iv) Ti mi chc ti bn, trc tin xy ra s tng hp cc on mi RNA (primer) bi v cc DNA polymerase t n khng th bt u tng hp mi c. Mt khc, do hai si n ca mi chc phn cc ngc chiu nhau trong khi cc enzyme DNA polymerase v RNA polymerase ch xc tc theo chiu 3' 5', cho nn phng thc ti bn DNA din ra trn hai si khun l khng ging nhau: mt si lin tc hay cn gi l si dn u (leading strand) v mt si khng lin tc hay cn gi l si ra chm (lagging strand). Kiu ti bn nh th gi l ti bn na gin on (semi-discontinuous). S tng hp DNA khng lin tc di dng cc on ngn do R. Okazaki pht hin u tin nm 1969; sau ny chng c gi l cc on Okazaki (Okazaki fragments). Sau , cc on mi s c ct b v ch trng c thay bng DNA, v cc on Okazaki c ni li bi enzyme DNA ligase.

2. Cc enzyme tham gia ti bn DNA Mc d mi nhm sinh vt c mt h thng cc enzyme v protein ti bn ring, song chng c cc enzyme vi vai tr chung nht sau y: (1) Protein nhn bit v bm vo khi im t hnh thnh nn "phc hp m" (open complex). E. coli, l protein dnaA. (2) DNA gyrase: m cun DNA siu xon pha trc mi chc ti bn. (3) Helicase: tho xon DNA si kp ti mi chc to thnh cc vng si n. E. coli, n cng gi l protein dnaB hay protein rep. (4) Protein SSB (single strand binding protein): bm vo cc vng DNA si n do helicase tch ra, gi cho tm thi khng dnh tr li v nh mi si n mi c th lm khun (template) cho ti bn. (5) Primase: tng hp RNA mi. E.coli, n cn gi l protein dnaG. (6) Cc DNA polymerase xc tc chnh cho vic tng hp DNA mi nh c hot tnh xc tc: polymerase 5'3', mt s cn c hot tnh c sa: exonuclease 3'5'. E. coli, l DNA polymerase III. (7) DNA polymerase va ct b dn on mi i trc nh hot tnh ct b exonuclease 5'3', va ko di on Okazaki theo sau lp ch trng. E. coli, l DNA polymerase I. (8) DNA ligase: hn lin khe h gia cc on DNA mi bng cch hnh thnh lin kt 3',5'-phosphodiester.
Bng 5.5 c tnh ca cc DNA polymerase E. coli v ngi DNA polymerase E. coli Polymerase 5'3' Exonuclease 3'5' Exonuclease 5'3' DNA polymerase ngi nh v Ti bn Sa cha Chc nng Polymerase 5'3' Exonuclease 3'5' Exonuclease 5'3' Primase Kt hp vi PCNA* Mu trt (Processivity) Tng hp si pol I c c c nhn c khng c khng khng c khng thp
ra chm
pol II c c khng nhn khng c c khng khng khng khng

sacha
pol III c c khng ty th c khng c c khng khng khng

c hai
nhn c c c c khng khng c cao

dn u
nhn c c c c khng khng c

ra chm

Lu : (i) E. coli, c ba loi protein dnaB, dnaC v dnaG hp thnh mt phc hp c tn l primasome; trong protein dnaC bm protein dnaB. Bng 5.5 m t cc c tnh ca cc DNA polymerase E. coli v ngi i din cho cc nhm tng ng, prokaryote v eukaryote. (ii) Tt c cc DNA polymerase u cn c mi vi mt nhm 3'-OH t do; xc tc tng hp chui theo mt hng 5'3'; v ch mt s enzyme ny c hot tnh c sa (proofreading activity) 3'5'. (iii) Ngc vi E. coli, cc t bo ngi c t nht nm loi DNA polymerase, trong ba loi chu trch nhim ti bn DNA nhn l cc DNA polymerase alpha, delta v epsilon. Polymerase chu trch nhim chnh cho tng hp trn si dn u (leading strand) v thm ch c si ra chm (lagging strand). Polymerase kt hp vi mt RNA primase v c coi l c cc hot tnh cho tng hp mt on mi RNA-DNA ngn. C iu khng chc chn lm khi ni v chc nng ca alpha trn si ra chm. V dng nh n thiu mt hot tnh c sa, nn khng th tng hp DNA vi chnh xc cao v nh vy c th n khng phi l polymerase chnh trong tng hp si ra chm. Polymerase c th hot ng nh DNA polymerase I ca vi khun. Loi khng nguyn trong nhn lm tng sinh t bo (proliferating cell nuclear antigen = PCNA) c vai tr trong c ti bn ln sa cha. Mt trong cc chc nng ca n l dng lm nhn t trt (processivity factor) cho DNA polymerase v . PCNA gi DNA polymerase vi si khun tng hp DNA nhanh. (iv) Ngoi ra cn c mt s enzyme v protein c th tham gia vo khu kt thc ti bn, tng hp cc u mt (telomere) hoc tham gia ct ni trong qu trnh ti t hp. 3. C ch ti bn DNA Trong phn ny, chng ta tm hiu k c ch ti bn vi khun E. coli, v nu mt t vn lin quan eukaryote m i din l DNA ngi. 3.1. Giai on khi u ca s ti bn (initiation of replication) i vi nhim sc th E. coli, s ti bn bt u ti mt khi im c th gi l Ori (hnh 5.18). Trong khi , trn mi nhim sc th eukaryote c nhiu khi im ti bn hai hng nh th (hnh 5.19). Qa trnh din bin ti khi im cho n lc hnh thnh hai chc c th tm tt (theo Kelman v O'Donnell, 1994) nh sau: (1) Cc protein bm khi im dnaA c tng hp bm Ori to ra cu trc nucleoprotein chuyn ho ca khi im; (2) Sau , cu trc ny m xon vng DNA giu AT hnh thnh "phc hp m" (open complex); v (3) Hai phn t helicase dnaB chui vo phc hp m lm m xon khi im theo c hai

hng, to thnh hai chc ti bn (replication fork). Khi c hai si n ca mi chc c tch ra th cc protein SSB bm vo. Nh vy cc si n c dng lm khun hiu qu cho cc enzyme ti bn hot ng. Khi u trong ti bn DNA l s tng hp mt on mi ngn bi primase, m E.coli l phc hp primasome (th m u) nh ni trn. Kch thc on mi cc sinh vt khc nhau l khng ging nhau, v thng khng vt qu 12 ribonucleotide. E. coli, theo kt qu nghin cu ca b Okazaki v cs (1985), on ny di 10-12 nucleotide.
Khi im ti bn
Cc u mt khng y
Hnh 5.19 Nhiu khi im ti bn (origins of replication) trn mt nhim sc th eukaryote. y cng cho thy cc u mt (telomeres) khng c tng hp y , c th l cc u 5'.
3.2. Giai an ko di (elongation) Mt khi primosome tng hp xong mt mi, l lc s ko di chui DNA mi bt u. E. coli, enzyme then cht cho qu trnh ny l DNA polymerase III hon chnh (holoenzyme), mt phc hp gm mi polypeptide khc nhau. Mi phn t cho mt si khun. S ko di trong sut qu trnh tng hp DNA E. coli r rng l cn ti mt replisome (th ti bn) do kt hp gia primasome v DNA polymerase III hon chnh. Tc tng hp trung bnh l 1.000 nucleotide mi giy. S ti bn DNA trn mi chc, nh ni trn, xy ra theo kiu na gin on (semi-discontinuous). C th qu trnh nh sau (hnh 5.20): Trn si khun dn u (3'5'): Trc tin, enzyme primase hay primasome ch tng hp mt on mi RNA vi u 3'-OH t do. Sau , enzyme hon chnh DNA polymerase III (replisome) bt u ko di chui DNA mi sinh trng theo chiu 5'3' mt cch lin tc.

Trn si khun ra chm (5'3'): S ko di din ra khng lin tc di dng cc on Okazaki. Kch thc trung bnh mi on Okazaki E. coli l 1.000 - 2.000 nucleotide; eukaryote l 100-200; v ni chung l 100-1.000 nucleotide. Qu trnh ny i hi s "mi ha" nhiu ln v c tnh chu k, vi s tham gia ln lt ca bn enzyme nh sau: (i) primase tng hp mt on mi RNA; (ii) DNA polymerase III hon chnh ko di on Okazaki; (iii) DNA polymerase I va ct b dn tng nucleotide ca on mi va lp khong trng bng cch ko di dn on Okazaki theo sau;
Hng ti bn chung
Si ra chm
Si dn u
Hnh 5.20 S ti bn na gin on trn mt chc.
(iv) DNA ligase hn lin khe h cn li gia hai on Okazaki k nhau bng mt lin kt 3',5'-phosphodiester. Qu trnh c din ra lun phin nh vy, v cui cng si DNA mi c tng hp tr nn di ra v lin tc. Cn eukaryote, vai tr ca cc enzyme v protein tham gia vo giai on ko di trn c hai si khun c phn tch trn. 3.3. Giai an kt thc (termination) Do c im cu trc nhim sc th hai nhm prokaryote v eukaryote l hon ton khc nhau, c bit l cu trc cc u mt nhim sc th eukaryote c pht hin gn y (hnh 5.21), nn c ch kt thc ti bn ca chng cng khc nhau. 3.3.1. Vn kt thc ti bn prokaryote hon thnh vic ti bn DNA, t bo phi lp y cc khong trng do cc RNA mi b ct b li. i vi cc DNA mch vng nh ca vi khun chng hn, khng c vn lp tt c cc khong trng bi v bao gi cng c u 3' DNA khc nm pha trc dng lm mi. Tht vy, c hai chc ti bn c bt u t mt khi im duy nht (ori), v di chuyn hu nh cng tc , theo hai hng i lp nhau xung quanh nhim sc th mch vng cho ti khi chng gp nhau ti mt im kt thc chung i din vi ori. Thc ra, theo Bastia v cs (1997) cng nh

nhiu tc gi khc, y l vng cha cc trnh t c th gi l cc im kt thc ti bn (replication termini). V ti cc trnh t c th ny c cc protein kt thc ti bn (replication terminator protein = RTP) bm vo v cc phc hp protein-DNA ny ngn cn s di chuyn ca cc chc ti bn theo mt cch phn cc hoc nh hng c th. S ngng li ca cc chc ti bn ti vng kt thc to nn bc u tin trong qu trnh hon thnh mt vng ti bn v sau , tch hai nhim sc th con ri ra mt cch c trt t nh xc tc ca topoisomerase IV. Lu : (1) Cc nghin cu gn y cho thy RTP ca E. coli c trng lng phn t ~36kD, c m ha bi gene tus (ter) v trong mi t bo c ~80 bn sao ca protein ny c duy tr hu nh n nh trong sut chu k t bo. Cn RTP ca Bacillus subtilis l mt dimer c trng lng phn t mi tiu n v l 14.500 kD, c m ha bi gene rtp. (2) Cc trnh t nht tr (consensus sequences) ca vng kt thc ti bn E. coli (R6K) v B. subtilis, theo Bastia v cs (1997), nh sau:
E. coli (R6K) 5'NN(A/T)(A/T)(A/T)G(A/T)(A/G)TGTTGTAACTA(A/C)NN3' B.subtilis 5'ACT(A/G)AN(T/A)(A/G)(A/C)(A/T)(C/T)(T/A)(A/G)TG(T/A)AC/CTTCAN3'
3.3.2. Vn kt thc ti bn eukaryote Nh chng ta u bit, mi nhim sc th eukaryote cha mt phn t DNA si kp mch thng kt hp vi nhiu loi protein, c cc u mt c trng gi l telomere. Mt khi an mi u tin trn mi si c loi b th n khng c cch no b p li khong trng , bi v DNA khng th no ni rng theo chiu 3'5', v cng chng c u 3'
(a)
(b)
Hnh 5.21 (a) nh chp cho thy cc u mt nhim sc th - telomeres c mu vng c trng, v (b) s minh ha t chc ca telomere ngi.
nm pha trc nh trong cc DNA mch vng. Nu qu thc nh vy th cc si DNA s ngn bt i sau mi ln ti bn. Vy cc t bo eukaryote

gii quyt vn ny nh th no?
Ko di
Chuyn dch
Ko di
Hnh 5.22 M hnh ca Carol Greider v c ch tng hp cc on lp telomere trn si giu G nh telomerase Tetrahymena. T y c th hnh dung cc bc tip theo: lp khong trng trn si i din giu C c thc hin bi primase v DNA polymerase, v cui cng l ct b on mi.
Cc kt qu nghin cu u tin ca Elizabeth Blackburn v cc ng s ca b gii p cho vn ny (hnh 5.22). Cc telomere khng cha cc gene; thay v th chng c cu trc n gin gm nhng trnh t ngn (6-8 cp base) lp li ni tip c ngn ln v c th cho tng loi. Chng hn, Tetrahymena, mt nhm ng vt nguyn sinh c lng t, n l (TTGGGG)n; Caenorhabditis: (CCCTCCC)n; bn Oxytrichia thuc Euplotes: (CCCCAAA)n... Cn cc ng vt c v v ngi, n l 5'-TTAGGG-3' c lp li khong 1.000-2.000 ln (theo Yakoob v cs 1999, khong bin thin pht hin c trong cc t bo cc giai on khc nhau l 150-2000 ln). Cc on lp ny c gn thm vo u 3' ca cc si DNA, khng phi bng ti bn bn bo ton, m bng mt enzyme gi l telomerase. Nu nh telomerase khng cho php mt c ch ti bn bn bo ton, th n khng th s dng mt si DNA lm

khun cho si kia. Blackburn cho thy rng tnh cht c th ny nm ngay trong chnh bn thn telomerase, v do mt RNA nh trong enzyme. Nh vy, telomerase l mt ribonucleprotein v l mt enzyme phin m ngc (reverse transcriptase), tng hp DNA t mt khun RNA. Chng hn, telomerase ca Tetrahymena c mt RNA di 160 nucleotide c cha trnh t 3'-CAACCCCAA-5' lm khun cho tng hp cc on lp 5'TTGGGG-3' trong cc telomere ca Tetrahymena (hnh 5.22); ngi, phn t RNA trong telomerase di khong 450 nucleotide c cha trnh t 3'-AAUCCC-5' (nm trong on ng vi v tr 46-56) lm khun cho tng hp cc on lp telomere 5'-TTAGGG-3' trn cc si n c u mt 3'. Sau , si i din, DNA polymerase c th hon thnh vic tng hp "cc u mt khng y " (incomplete ends) trn si i din sau khi c mi ha bn trong cc telomere ; v cui cng on mi s b ct b v khe h c ni li. Cc nghin cu gn y cho thy rng: enzyme telomerase ch c mt trong cc t bo mm (germ cells), k c cc t bo gc ca phi (embryonic stem cells); cc t bo ung th (cancer cells); v cc eukaryote n bo nh Tetrahymena thermophila. Trong khi , cc t bo soma bnh thng ca ng vt c v khng thy c telomerase. iu cho php l gii ti sao cc t bo mm cng nh cc t bo ung th c kh nng phn chia gn nh l v hn; hay ni cch khc, telomerase v s duy tr chiu di telomere chnh l cha kha cho s bt t ca t bo. Ngc li, hu qu ca s vng bng telomerase trong cc t bo soma (do gene m ha n bt hot) l ch: c sau mi ln nguyn phn (mitosis), tt c 92 telomere ca cc t bo soma ngi ng lot mi ci b mt i chng 100 cp base, ngha l khong 16 on lp TTAGGG. Theo l thuyt, vi tc ny th sau 125 ln nguyn phn cc telomere s bin mt hon ton. Trn thc t, cc th nghim khc nhau k t khm ph u tin ca Leonard Hayflick vo u thp nin 1960 cho n nay (Greider v Blackburn 1996; Hayflick 1997; Shay v cs 2004; ...) xc nhn rng: ch sau khong 30-50 ln nguyn phn, cc t bo soma mt hn kh nng phn chia v i vo giai on lo ha (senescence) v cht t nhin. Ci c gi l gii hn Hayflick (Hayflick limit). Chnh qu trnh rt ngn telomere theo thi gian (ngha l s ln nguyn phn) nh th khin ngi ta lin tng ti s tn ti ca ci gi l "ng h di truyn cho s lo ha" (genetic clock for aging), v bt u tin hnh cc nghin cu v "hiu ng v tr telomere" (telomere position effect = TPE) (Borek 2002). l l do ti sao cc t bo soma c s ln phn chia hu hn trc khi chng cht v cng l l do ti sao tt c

chng ta cng nh mi sinh vt u phi cht. Nh th, ci cht tt yu trn phng din sinh hc, ci cht t nhin ca thn xc c l gii kh r rng v y . V t cc nghin cu ny m ra trin vng to ln cho liu php ung th (cancer therapy) cng nh vn lo ha v bnh tt pht sinh t con ngi (Greider v Blackburn 1996;Yakoob v cs 1999; Borek 2002; Shay v cs 1998, 2004).
VI. Ti bn ca cc b gene RNA (RNA genomes)

1. c im ti bn ca cc b gene RNA virus v hu qu ca chng cc virus c b gene RNA, phng thc ti bn ca chng r rng l khc vi cc h thng ti bn DNA bit. Trc ht, cc enzyme tng hp RNA khng cn c mi (primer), v vy khng c c hi cho vic c sa (proofreading). Ngoi ra, v mt di truyn RNA l mt phn t km bn vng so vi DNA bi v nhm hydroxyl c mt nguyn t C2' ca gc ng cho php ct t (thy phn) lin kt phosphodiester gia hai ribonucleotide v to thnh dng phosphodiester vng gia cc nhm hydroxyl ca C2' v C3' trong cng mt gc ng; sau cu trc vng ny m ra v li nhm phosphate v tr C3'. S kt hp ca hai c im trn l nguyn nhn lm hn ch kch thc cc b gene RNA. Cc b gene ny khng th sinh trng qu ln, hoc khng th trnh khi t l sai st cao trong qu trnh ti bn (10-3-10-4). V vy, trn thc t, b gene RNA ln nht c bit l mt RNA si n vi chng 29.600 base. l trng hp ca virus gy bnh st vim phi cp (SARS) thuc h coronavirus c pht hin hi thng 3/2002 ti mt s nc chu , Canada... cng l l do ti sao cc virus RNA cho nhiu bin th khc nhau n nh vy, mc d kch thc b gene u c ln. iu ny lm cho cc virus RNA thc s tr thnh mi him ha bi v chng c th tin ha nhanh xm nhp vo cc h thng min dch ca vt ch. Chng hn, cc b gene HIV c nhiu bin th gy kh khn cho vic bo ch cc vaccine v thuc chng li tt c cc bin th ca HIV. Di y nu khi qut v hai kiu ti bn ca cc virus RNA. 2.Ti bn ca b gene RNA Kiu truyn thng tin trc tip t RNA sang RNA xy ra trong cc t bo ly nhim virus RNA nh: virus m thuc l v nhiu virus thc vt khc, k c cc phage RNA nh MS2... B gene RNA ca cc virus ny c mang gen m ho enzyme ti bn c th. Sau khi c tng hp trong t bo ch, enzyme ny s dng bn thn RNA virus lm khun tng hp cc phn t RNA b sung. n lt mnh cc RNA li lm khun tng hp tr li cc phn t RNA ca cc virus th h con .

3. Phin m ngc Phin m ngc (reverse transcription) l kiu truyn thng tin t RNA sang DNA, ch xy ra trong cc t bo ng vt v ngi b ly nhim bi mt s virus mang mt si RNA c kh nng gy khi u hoc hai si RNA nh trng hp HIV chng hn. Trn mi si RNA li ca cc virus ny c mang mt enzyme phin m ngc (reverse transcriptase). Khi xm nhp vo t bo ch, enzyme ny s dng RNA ca virus lm khun tng hp si DNA b sung (complementary DNA = cDNA). Sau , si cDNA ny c th lm khun tng hp tr li b gene ca virus (cDNARNA), hoc tng hp ra si DNA th hai b sung vi n (cDNADNA) nh trong trng hp virus gy khi u m kt qu l to ra mt cDNA si kp. Phn t DNA si kp c tng hp trc tin trong qu trnh ly nhim c th xen vo DNA ca vt ch v trng thi tin virus (provirus). V vy, provirus c truyn li cho cc t bo con thng qua s ti bn ca DNA vt ch, ngha l cc t bo con chu ca vt ch cng b chuyn sang tnh trng c mm bnh ung th. Cc t bo ung th ny mt kh nng kim sot s sinh trng - phn chia in hnh ca t bo bnh thng; chng tng sinh rt nhanh v to ra khi u (tumor). chnh l c ch gy ung th bi virus. Ngy nay, ngi ta c th tinh chit cc enzyme phin m ngc phc v cho k thut to dng cDNA ti t hp (chng 10).
Cu hi v Bi tp
1. Trn c s cc c im ca m hnh Watson-Crick, hy cho bit: (a) M hnh ny cho php gii thch cc quy lut Chargaff nh th no? (b) C s ca c tnh i song song trong chui xon kp DNA l g? (c) M hnh ny gi ln kh nng t ti bn ca DNA ra sao? 2. Hy xc nh hm tng i (%) ca cc nucleotide trong DNA ca ngi, c hi (Salmo salar) v g (Gallus domesticus). Bit rng t l (A+T)/(G+C) tng ng ca cc loi ny l 1,52; 1,43; v 1,34. 3. Da vo cu trc ha hc lp th ca cc base (A,T,G v C), hy gii thch: (a) Ti sao trong ADN ch tn ti hai kiu kt cp A-T v G-C m khng c cc kiu khc? (b) Cc kiu kt cp A-C v G-T c th xy ra khi no v gy ra hu qu g? Gii thch v cho cc s minh ha. 4. Gi s t l (A+T)/(G+C) mt si ca chui xon kp ADN l 0,25. (a) Hy cho bit t l ny trn si b sung v trn c phn t? (b) Nu cho rng 0,25 l ca t l (A+G)/(T+C), th t l ny trn si b sung

v trn c phn t s nh th no? 5. Hy ch ra nhng im ging nhau v khc nhau gia prokaryote v eukaryote v cc vn sau: (a) Khi im ti bn; (b) Cc enzyme tham gia ti bn; v (c) Cc c ch m u, ko di, v kt thc ti bn. 6. Da vo hiu bit v thuyt di truyn nhim sc th, hy gii thch th nghim Griffith v s bin np ph cu khun S. pneumoniae. 7. So snh cu trc ca cc nucleotide v cc chui polynucleotide cng nh cu trc phn t ca DNA v RNA. 8. Th no l bin tnh v hi tnh ca DNA? Cc hin tng ny c ngha nh th no i vi cc b gene sinh vt v k thut di truyn? 9. Th no l gi tr C (C-value) v nghch l gi tr C (C-value paradox)? Cho th d v nu ngha v mt l lun ca hiu bit ny. 10. S ti bn ca cc b gene RNA mt s virus khc vi cc h thng ti bn DNA nhng im no? Ti sao kch thc cc b gene RNA thng nh, nhng chng li c kh nng gy nguy him thc s i vi cc vt ch? Hy gii thch mt c ch gy ung th bi virus.
Ti liu Tham kho

Ting Vit H Hunh Thy Dng. 1997. Sinh hc Phn t. NXB Gio Dc. Phm Thnh H. 2000. Di truyn hc. Ti bn ln II, NXB Gio Dc. Phan C Nhn. 2001. Di truyn hc ng vt. NXB Khoa hc v K thut, H Ni. Phan C Nhn. 1999. Cng ngh ADN ti t hp. Trong: Di truyn hc tp II (Phan C Nhn, ch bin). Trang: 257-303. NXB Gio Dc, H Ni. Hong Trng Phn. 1995. Mt s vn v Di truyn hc hin i (Ti liu BDTX cho gio vin THPT chu k 1993-1996). Trng HSP Hu. Hong Trng Phn. 1997. Di truyn hc Phn t. NXB Gio Dc. Watson JD. 1968. Chui xon kp: Hi k v vic pht minh ra cu trc ca DNA. (Bn dch ca L nh Lng v Thi Don Tnh). NXB Khoa hc v K thut, H Ni, 1984. Ting Anh Alberts B, Johnson A, Lewis J, Raff M, Robert K, Walter P. 2002. Molecular Biology of the Cell. Garland Science, NY. Bastia D, Manna AC, Sahoo T. 1997. Termination of DNA replication in

prokaryotic chromosomes. In: Genetic Engineering, Vol. 19. (Setlow JK ed.) pp 101-119. Plenum Press, New York, USA. Borek C. 2002. Telomere Control and Cellular Aging: http://www.lef.org/magazine/mag2002/oct2002 report telo 01.html. Donovan S and Diffley JFX. 1995. Replication origins in eukaryotes. In: Current Opinion in Genetics & Development (Herr W and Kingston R, eds.),Vol.5(2): 203-207. Current Biology Ltd, UK. Greider CW and Blackburn EH. 1996. Telomere, Telomerase and Cancer. Scientific American, 2/96, p.92: http://www.genethik.de/telomerase.htm. Hamlin JL and Dijkwel PA. 1995. On the nature of replication origins in higher eukaryotes. In: Current Opinion in Genetics & Development (Herr W and Kingston R, eds.),Vol.5(2): 153-161. Current Biology Ltd, UK. Harley CB, Villeponteau B. 1995. Telomeres and telomerase in aging and cancer. In: Current Opinion in Genetics & Development (Herr W and Kingston R, eds.),Vol.5(2): 249-255. Current Biology Ltd, UK. Hayflick L. 1997. Mortality and Immortality at the Cellular Level. A Review. Copyright 2005BioProtNetwork:webmaster@ protein.bio.msu.su. Kelman Z, O'Donell M. 1994. DNA replication: enzymology and mechanisms. In: Current Opinion in Genetics & Development (Stillman B and Green M, eds.),Vol.4(2): 185-195. Current Biology Ltd, UK. Lingner J and Cesh TR. 1998. Telomerase and chromosome end maintenance. In: Current Opinion in Genetics & Development (Allis CD and Gasser SM, eds.),Vol.8(2): 226-232. Current Biology Ltd, UK. Lewis R. 2003. Human Genetics: Concepts and Applications. 5th ed, McGraw-Hill, Inc, NY. Price CM. 1999. Telomeres and telomerase: broad effects on cell growth. In: Current Opinion in Genetics & Development (Kadonaga JT and Grunstein M, eds.),Vol.8(2): 218-224. Current Biology Ltd, UK. Russell PJ. 2003. Essential Genetics. Benjamin/Cummings Publishing Company, Inc, Menlo Park, CA. Shay JW and Wright WE. 1998. Teleomerase, Cellular Senescence and Cancer. Page maintained by: Cell Biology and Neuroscience. Shay JW. 2002. Telomeres, Aging, and Tumorigenesis. Gac Md Mx Vol.138 Suplemento No.1, 2002, pp.S2-S3. Shay J, Schneider E, Masoro EJ. 2004. Is there a genetic clock for aging?: www.infoaging.org/b-tel-home.html. Tamarin RH. 1999. Principles of Genetics. 6th ed, McGraw-Hill, Inc, NY.

Twyman RM. 1998. Advanced Molecular Biology. BIOS Scientific Publishers Ltd/ Springer-Verlag Singapore Pte Ltd. Watson JD, Hopkins NH, Roberts JW, Steitz JA, Weiner AM. 1987. Molecular Biology of the Gene. 4th ed, Benjamin/Cummings Publishing Company, Inc, Menlo Park, CA. Weaver RF, Hedrick PW. 1997. Genetics. 3rd ed, McGraw-Hill Companies, Inc. Wm.C.Browm Publishers, Dubuque, IA. Yakoob J, Hu G-L, Fan X-G, Zhang Z. 1999. Telomere, telomerase and digestive cancer. World Journal of Gastroenterology, 5(4):334-337. Mt s trang web http://www.lef.org/magazine/mag2002/oct2002 report telo 01.html. http://www.genethik.de/telomerase.htm. http://www.infoaging.org/b-tel-home.html. http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/ http://www.ornl.gov/hgmis http://www.horizonpress.com/pcr/ http://dir.niehs.nih-gov/dirlmg/repl.html

Chng 6
Gene v Qu trnh Sinh tng hp Protein

Gene, n v thng tin c truyn t cha m cho con ci, l khi nim then cht ca di truyn hc. Ni n gene tc l ni n DNA v cc quan h ca n vi RNA v protein di dng s sau y, c gi l L thuyt trung tm (Central Dogma) ca Sinh hc Phn t (hnh 6.1). Trong cc si n ca DNA c dng lm khun cho ti bn (replication; nh xt Ti bn chng 5). Mt khc, tng an xc nh ca n (tc cc gene) c Phin m th lm khun cho s tng hp cc RNA trong mt qu trnh gi l phin m (transcription). n Dch m lt, cc phn t RNA ny li lm khun cho s tng hp cc chui polypeptide m t to thnh cc protein; qu trnh ny c
Hnh 6.1 L thuyt trung tm ca Sinh hc Phn t
gi l dch m (translation), bi v n chuyn bc thng tin di dng cc nucleotide thnh ra sn phm c xy dng bng cc amino acid. Hai qu trnh sau c coi l hai giai an chnh trong s biu hin ca gene m ha protein (protein coding gene). Thc ra, s biu hin ca mt gene chu s kim sot nhiu cp khc nhau (chng 7). Trong chng ny, chng ta s ln lt tm hiu s pht trin ca khi nim gene, cu trc v chc nng protein - sn phm ca gene, bn cht ca m di truyn, v cc qu trnh phin m v dch m.
I. S pht trin ca khi nim gene

1. Cc quan nim ca Mendel v Morgan v gene Mendel l ngi u tin nu ln nh ngha v gene nm 1865 (thut ng ny c Johannsen a ra nm 1909). Theo , gene l n v di truyn tn ti dng ht ring bit, xc nh mt tnh trng c th trong cp tnh trng tng phn. y mi ch l s suy lun thun ty, khng c c s vt cht c th. Quan nim chnh xc hn v c s vt cht v chc nng ca gene ny

sinh t nhiu ngun nghin cu c lp nhau trong sut 50 nm u ca th k XX. Trng phi Morgan sau khi xc nh cc gene nm trn nhim sc th v xut phng php lp bn gene bng ti t hp, khng nh rng cc gene l nhng n v c s v khng chia nh ca vt cht di truyn v c cu trc ln chc nng; chng lin kt vi nhau theo kiu thng hng trn nhim sc th. 2. Gi thuyt mt gene - mt enzyme ca Beadle v Tatum Mt hng nghin cu khc tp trung vo phng din chc nng sinh ha ca gene. Nm 1902, Archibald Garrod gi rng ri ln chuyn ha alkapton niu (alcaptonuria) bt ngun t mt sai hng ca mt enzyme c th v c di truyn theo kiu ln nhim sc th thng, m ng gi l sai st chuyn ha bm sinh. n nm 1941, Beadle v Tatum mi lm sng t tng trn bng cc th nghim gy t bin bng tia X Neurosporora. gii thch cc tn thng sinh ha c th do t bin, h xut "gi thuyt mt gene - mt enzyme" ni ting; n c xem nh l m hnh v chc nng ca gene, m ng cho s ra i ca di truyn sinh ha. V sau, quan nim "mt gene - mt enzyme" c m rng thnh "mt gene - mt protein", v tip tc chnh xc ha bng mnh "mt gene - mt polypeptide". Tht vy, t khi Avery v cc ng s chng minh DNA l vt cht mang thng tin di truyn vo nm 1944, v c bit l sau khi Watson v Crick khm ph ra cu trc phn t DNA nm 1953, quan nim v gene khng ngng c pht trin v chnh xc ha. V mt cu trc, gene l mt on xc nh ca b gene (DNA hu ht sinh vt v RNA mt vi virus). V phng din chc nng, nh chng ta r, khng phi mi gene u m ha cc enzyme m mt s m ha cc polypeptide vi cc chc nng khc nhau, v mt s m ha cc phn t RNA chc nng nh RNA ribosome (rRNA) v RNA vn chuyn (tRNA). Hn na, thng tin trong gene c th c s dng mt cch c chn lc sinh ra nhiu hn mt loi sn phm (cc gene phn on). Trc tin, ta hy tm hiu cng trnh nghin cu ca Benzer v cu trc tinh vi ca gene. 3. Quan nim ca Benzer v cc n v cu trc v chc nng di truyn Cc cng trnh nghin cu ca Seymour Benzer (t 1957 n 1961) v ti t hp phage T4 cho thy rng, gene theo quan nim ca Morgan c th chia nh thnh cc n v nh hn. ng a ra cc thut ng muton, recon v cistron nh ngha cc n v khng chia nh tng ng l t bin, ti t hp v chc nng. Bng cch lai cc th t bin ca cng mt gene c ngun gc c lp nhau trong khi cho ly nhim phage, lm xut hin phage kiu di. iu ny ch c th xy ra bi s

ti t hp bn trong gene (intragenic recombination), nu nh cc phn nh ring bit ca gene u b t bin. iu ny chng t rng gene b phn chia thnh cc n v nh hn thng qua ti t hp v dt bin. Tuy nhin, v kch thc ca muton v recon c coi l tng ng vi mt cp nucleotide, cho nn ngy nay t thn hai n v ny khng cn gi tr s dng na. Thut ng cistron ca Benzer c ngha l n v chc nng di truyn khng chia nh. iu ny c th xc nh bng s phn tch b sung (complementation analysis), trong gene m c th l sn phm ca n c trc nghim v kh nng b p cho mt t bin ti mt gene tng ng trong cng t bo. S b sung lin tip lm phc hi kiu hnh di.
cistron 1 cistron 2 cistron 1 cistron 2

S I
Kiu di S
X X
P Kiu di
(a)
cistron 1 cistron 2
(b)
cistron 1 X
S
X
S I
P Kiu di bin
cistron 2
Th t
(c)
(d) X
Hnh 6.2 S minh ha trc nghim cis-trans: (a) con ng chuyn ha bnh thng; (b) trc nghim cis; (c) v (d) trc nghim trans. Ch thch: S-c cht (subtrate); I- sn phm trung gian (intermediate); P- sn phm cui cng (product), y l sc t c trng cho kiu hnh di; cc mi tn () ch cc enzyme sn phm sinh ra t cc cistron 1 v cistron 2.
C s ca phn tch b sung l trc nghim cis-trans (cis-trans test), m t y ny sinh ra thut ng cistron, trong cc cp t bin bt ngun c lp c xt cc cu hnh cis (u) v trans (lch). Trc nghim cis c dng lm i chng, v nu nh c hai t bin u c mt trong mt b gene th b gene kia phi l kiu di c hai locus v sinh ra cc sn phm gene bnh thng, do cho ra kiu hnh di (hnh 6.2b). Trc nghim trans l php th b sung v xc nh gi hn ca n v chc nng. Nu nh cc t bin nm trong cc gene khc nhau, khi chng c mt cu hnh trans, mi mt b gene c th b sung sn phm

m gene kia khng to ra c. Khi c tt c cc sn phm gene cn thit th t bo l kiu di (hnh 6.2c), ngha l c s b sung dng tnh (positive complementation). Nu nh c hai t bin thuc cng mt gene, khi chng c mt cu hnh trans, th mi mt b gene c th mang mt bn sao t bin ca gene v khng c sn phm hot ng chc nng c to ra trong t bo, ngha l khng c s b sung (hnh 6.2d). S phn tch b sung vi khun v nm men bia cng ch ra rng gene l mt cistron, ngha l gene c nh ngha nh l mt n v chc nng. Phng php ny t ra hu ch cho vic khng nh chc nng ca gene, xc nh s lng cng nh trt t hot ng ca cc gene trong mt con ng chuyn ha no . Vy cistron l g? Cistron l mt on xc nh ca DNA (hay b gene ni chung) mang thng tin cu trc ca mt polypeptide c th m gii hn ca n c xc nh bng trc nghim cis-trans. Kch thc trung bnh ca mt cistron l 1.200 cp base. Nh vy, cistron chnh l gene cu trc theo ngha hp hay gene m ha protein. Mt cch tng i, theo ngha rng, c th nh ngha gene l mt an xc nh ca b gene m ha thng tin ca mt polypeptid hoc mt phn t RNA chc nng (nh tRNA, rRNA...). Tuy vy, nh ngha ny khng th bao gm y chc nng v cu trc ca gene trong ton b sinh gii, bi v cc chin lc cho s biu hn gene v t chc b gene cc vi khun v eukaryote l rt khc nhau. 4. Mi quan h gene - cistron cc prokaryote v eukaryote 4.1. S tng ng gene - cistron cc b gene n gin cc prokaryote v eukaryote bc thp, thng c mt mi quan h n gin gia gene v sn phm ca n.Trong hu ht trng hp, c mt s tng ng mt gene - mt sn phm, v s ng tuyn tnh gia gene v chui polypeptide ca n c Ch.Yanofsky xc nhn nm 1961. V vy cc sinh vt ny, gene v cistron l tng ng: gene l n v chc nng di truyn, mang thng tin di truyn c biu hin trn vn.
(a)
(b)
Hnh 6.3 (a) vi khun, cc gene thng c sp xp trong mt operon v c phin m thnh mt phn t mRNA a cistron. (b) eukaryote, cc gene tn ti ring bit di dng n cistron.

Cng cn lu rng, vi khun, cc gene ng ngha vi vng m ha hay khung c m (open reading frame = ORF), trong khi cc eukaryote n ng ngha vi n v phin m (transcription unit). l do cc gene vi khun thng c sp xp trong mt operon (chng 7), v th c nhiu sn phm c dch m t mt mRNA a cistron (polycistronic mRNA; hnh 6.3a). Tri li, cc eukaryote, hu ht cc gene c phin m di dng mRNA n cistron (monocistronic mRNA; hnh 6.3b). 4.2. S khng tng ng gene - cistron cc b gene phc tp cc b gene eukaryote bc cao, thng thng c mt mi quan h phc tp gia gene v sn phm ca n (hnh 6.4). Hu ht cc gene ca eukaryote bc cao u c cha cc intron (intervening sequences), tc cc on khng m ha protein, nm xen gia cc exon (expressed sequences), cc on m ha protein. Cc gene nh vy c gi l gene phn on (split gene) hay gene t qung (interrupted gene); n c pht hin ln u tin bi Phillip Sharp vo nm 1977 (hnh 6.4).
(a) (b) (c) Hnh 6.4 nh hin vi in t v hnh m phng vic s dng vt d mRNA t bo cht c nh du pht hin gene phn on (a v b); m t vn tt s tng hp pre-mRNA v ct b cc intron to ra mRNA trng thnh t mt gene phn on c cha hai intron (c).
Cc khm ph v sau ny cn cho thy nhng s kin rc ri ny sinh trong cc gene phn on ny, ch: thng tin trong gene c s dng mt cch chn lc sinh ra nhiu sn phm khc nhau, gi l ct-ni c chn lc (alternative splicing) v.v. Cc sn phm c quan h v cu trc (v d, calcitonin/CGRP) thng c cc chc nng khc nhau. V l , cistron i khi c xem l tng ng vi exon ca gene eukaryote, v gene phn on c xem nh l mt chui cc cistron gi nhau (Twyman 1998). Ngoi ra, c vi trng hp trong cn ti hai gene sinh ra mt sn phm mRNA n thng qua kiu ct-ni cho (trans-splicing)

hoc bin tp RNA (RNA editing). Chng hn, khm ph mi nht cho thy glucose 6-phosphate dehydrogenase l mt enzyme c mt trong cc t bo hng cu ngi; n bao gm hai dng nh/th yu v ln/chnh yu (minor and major form); dng u c trnh t cc amino acid thuc gene trn nhim sc th X; v dng sau gm hai peptide c m ha t thng tin ca hai nhim sc th, cc amino acid trong on 1-53 c m ha trn nhim sc th s 6 v cc amino acid on tip theo 54-479 c m ha trn nhim sc th X (theo McClean 1998). Tt c nhng trng hp ny ni ln mt iu rng, a ra mt nh ngha chnh xc v gene khng h n gin t no. Tuy nhin, nhn ton cc th mt khi nim thng nht ni bt l, tt c cc sinh vt t vi khun E. coli cho n con ngi u c chung h thng mt m di truyn v c chung phng thc 'chuyn ti' thng tin trong gene thnh ra protein. 4.3. Cc thnh phn cu trc hay l t chc ca mt gene T chc ca mt gene c th bao gm cc vng ring bit vi cc chc nng c th (Bng 6.1; theo Twyman 1998, c sa i).
Bng 6.1 Cc thut ng c dng ch cc phn chc nng ca cc gene Thut ng Allele Cistron Vng m ha, khung c m (ORF) Gene phn on Gene nh ngha Mt bin th v trnh t ca mt gene (hoc marker di truyn khc, v d: trnh t RFLP, VNTR). Mt n v chc nng di truyn, mt vng ca DNA m ha mt sn phm c th. Mt vng ca DNA c dch m thnh protein. vi khun, l m gene. eukaryote, vng m ha c th b gin an bi cc intron. Mt gene m ha protein gm cc on khng m ha (intron) nm xen k gia cc om m ha (exon). vi khun, mt n v chc nng di truyn m ha hoc l 1 polypeptide ring hoc phn t RNA. eukaryote, 1 n v phin m c th m ha 1 hay nhiu sn phm hoc ng gp vo 1 sn phm. V tr ca mt gene trn mt nhim sc th, k c cc yu t iu ha k bn. Thut ng locus c dng theo cch ring ch v tr ca gene-marker bt k, yu t iu ha, khi im ti bn, v.v.. Mt locus ca vi khun c cha nhiu gene (m c phin m nh l mt bn sao polycistron n) v cc yu t iu ha chung ca chng. Mt trnh t khng hot ng chc nng vn c cu trc tng t mt gene hot ng chc nng. Bt k phn no ca n v phin m ca 1 gene RNA
Locus gene
Operon Pseudogene on m c

phin m n v phin m, vng c phin m Vng khng c dch m (UTR) Gene b phn chia (divided gene) hay operon gene RNA s b loi b trong khi to ra cc phn t RNA trng thnh. Mt vng ca DNA c phin m thnh RNA. cc eukaryote, l mt gene. vi khun, n c th bao qut nhiu gene. Bt k phn no ca n v phin m m khng c dch thnh protein. Cc UTR k bn mt vng m ha hay operon c gi l cc UTR 5' v 3'. Mt gene phn an vi cc exon cc locus ring bit c phin m ring r v khu ni li bi s ct-ni cho. Thc ra l cch gi sai v mi locus ng ra phi c coi nh l 1 gene ring.
bt k locus no, mt vng DNA c phin m c th gi l mt n v phin m (transcription unit). Nh cp, prokaryote, mt n v phin m c th gm nhiu gene; trong khi eukaryote, cc n v phin m hu nh bao gi cng tng ng vi mt gene n.
Hnh 6.5 Cu trc in hnh ca mt gene eukaryote.
i vi cc gene m ha protein, r rng l c mt s tch bit gia vng c dch m thnh chui polypeptide v vng khng c dch m. vi khun, vng c dch m l khung c m (ORF), trong cc gene phn cch nhau bng cc on m (spacer) c gi l cc vng khng m ha bn trong (internal noncoding region). Cc gene nm hai u ca mt operon cng c km bi mt vng khng m ha c tn l vng khng dch m 5' (5' untranslated region = 5' UTR) hay on dn u (leader sequence) v vng khng c dch m 3' (3' UTR) hay on ko sau (trailer sequence). V bn cht, chng l cc on iu ha; chng hn, vng 5'UTR kim sot s bm vo ca ribosome, cn vng 3'UTR thng ng vai tr quan trng trong s n nh mRNA. cc gene eukaryote, vng m ha cng c km bi cc vng UTR iu ha, v c hai vng UTR 5' v 3' cng nh khung c m c th b gin on bi cc on khng m ha (tc cc intron) m chng s c ct b trc khi xut mRNA trng thnh ra khi nhn. Nh th, bt k on no m rt cuc b loi b khi pre-RNA th c gi l cc on m c phin m (transcribed spacer). Cu trc in hnh ca cc gene m ha protein prokaryote v eukaryote c ch ra tng ng hnh 6.3a v hnh 6.5.

II. Cu trc v chc nng ca protein
1. Cu trc protein Cc protein l nhng polymer sinh hc c cu to bng cc amino acid ni kt vi nhau bng cc lin kt peptide. C 20 loi L--amino acid c pht hin trong cc protein ca cc t bo (hnh 6.6). V cu trc, ni chung, mi amino acid gm c mt nguyn t carbon alpha (C) v tr
A
Hnh 6.6 Hai mi loi amino acid pht hin c trong cc protein, vi bn nhm: A. Cc amino acid c chui bn tch in dng (3 bn tri) v m (2 bn phi); B. Cc amino acid c chui bn khng tch in; C. Cc trng hp c bit; v D. Cc amino acid c chui bn k nc.

trung tm, nh xung quanh n l mt nhm amin (-NH2), mt nhm carboxyl (-COOH), mt nguyn t hydro (-H) v mt gc R hay chui bn c trng cho tng loi amino acid (hnh 6.7a). Khi trng thi dung dch, cc nhm amin v carboxyl thng phn ly thnh trng thi ion, tng ng l +H3N- v -COO. Hai amino acid ni vi nhau bng mt lin kt peptide (CN) gia nhm carboxyl ca amino acid ny vi nhm amin ca amino acid k tip v loi tr mt phn t nc; c nh th cc amino acid kt ni vi nhau to thnh mt chui gm nhiu amino acid, thng c gi l polypeptide (hnh 6.7b). Mi chui polypeptide lun lun c chiu xc nh +H3N COO (do tc dng ca enzyme peptydyltransferase) v c c trng v s lng, thnh phn v ch yu l trnh t sp xp ca cc amino acid (hay cn gi l cu trc s cp, cu trc quan trng nht ca tt c cc protein do gene quy nh).
(a)
(b)
Hnh 6.7 (a) Cu trc chung ca mt amino acid; (b) s hnh thnh chui polypeptide, cu trc s cp ca tt c cc protein.
C bn mc cu trc ca cc protein c trnh by hnh 6.8. Trt t sp xp thng hng ca cc amino acid to thnh cu trc bc I (primary structure) ca protein. Cch thc cc amino acid ny tng tc vi cc amino acid ln cn bng cc mi lin kt hydro hnh thnh nn cu trc bc II (secondary structure) ca protein; hai dng ph bin ca cu trc bc II l: chui xon alpha (-helix) v tm beta (-pleated sheet). Cn hnh dng khng gian ba chiu ca mt chui polypeptide chnh l cu trc bc III (tertiary structure) ca n; hu ht cc protein u ly dng ny m ta gi l hnh cu (globular). V nhiu protein c cu trc gm hai hoc nhiu polypeptid cng hp nht trong mt protein phc tp, gi l cu trc bc IV (quaternary structure). y l mc cu trc cao nht ca protein; chng thng cha nhiu vng cu trc cun cht gi l cc domain, nh trong hemoglobin hoc cc khng th (xem hnh 6.9). 2. Chc nng protein Ni chung, protein l cc hp cht hu c lm nn s sng vi nhng chc nng thit yu khc nhau sau y: (i) Cc protein l thnh phn cu to c s ca cc t bo, bao gm

cc mng t bo, cc bo quan, b my di truyn ca chng. cng l cc protein dng si lm thnh cc c quan b phn trn c th cc ng vt, nh: collagen lm nn xng, sn, gn v da; keratin cu to nn cc lp ngoi cng ca da v tc, mng, sng v lng; (ii) Cc enzyme ng vai tr xc tc cho tt c cc phn ng ha hc trong t bo v c th u l nhng protein hnh cu. Quan trng nht l cc enzyme tham gia vo cc con ng chuyn ha v cc enzyme tham gia vo cc qu trnh truyn thng tin di truyn trong t bo.
Cu trc protein bc I l trnh t sp xp ca cc amino acid trong mt chui polypeptide. y l bc cu trc c s quan trng nht ca tt c cc protein do gene trc tip quy nh. Cc amino acid Tm beta Xon alpha Cu trc protein bc II xy ra khi trnh t cc amino acid trong mt chui polypeptide ni vi nhau bng cc lin kt hydro. Cu trc ny c hai kiu c bn, l: chui xon alpha (theo chiu xon tri) v tm beta (dng gp np). dng tm beta, hai chui polypeptide i song song xp cnh nhau; in hnh l cc si t. Tm beta Cu trc protein bc III xy ra khi cc lc hp dn no c mt gia cc vng xon Xon alpha v cc tm beta gp np trong mt chui alpha polypeptide, hnh thnh nn mt cu trc cun gp c dng khi cu. Mt s protein chc nng c cu trc kiu ny, nh myoglobin... Cu trc protein bc IV l mt protein gm hai hoc nhiu chui popeptide cng loi hoc khc loi kt hp vi nhau. C kh nhiu protein chc nng c kiu cu trc ny; mt s nh hemoglobin v chlorophyll, trong thnh phn cn c ion tng ng l Fe++ v Mg++. Hnh 6.8 Bn bc cu trc ca protein.
Hnh 6.9 Cu trc bc IV in hnh ca hemoglobin, tubulin v immunoglobulin. y cho thy s chui polypeptide v c bit l cc vng chc nng c trng ca hemoglobin v khng th immunoglobulin kiu IgG.
(iii) Cc hormone protein bt ngun t cc tuyn ni tit th khng hot ng nh cc enzyme. Thay cho s kch thch cc c quan ch, chng li khi u v kim sot cc hot ng quan trng, v d nh tc chuyn ha v sn xut ra cc enzyme tiu ha v sa. Insulin c tit ra t cc o Langerhans tuyn ty, iu ha s chuyn ha carbonhydrate bng cch kim sot cc mc glucose trong mu. Thyroglobulin (t tuyn gip) iu ha cc qu trnh chuyn ha ni chung; calcitonin cng t thyroid lm h thp mc calcium trong mu v.v. (iv) Cc khng th (antibodies) trong h thng min dch, cn gi l cc immunoglobulin, lm ra hng ngn protein khc nhau vn c sinh ra trong huyt thanh mu phn ng li vi cc khng nguyn (antigens). Chng ng vai tr bo v c th chng li s xm nhp ca cc vt l. (v) Ngoi ra, cc protein cn l ngun dinh dng chnh cung cp nng lng cho t bo v c th duy tr cc hot ng trao i cht v ln ln; cc protein nh hemoglobin mang cc sinh cht theo mu i khp c th; cc fibrinogen v fibrin c bin i t n vn c trong mu cn thit cho qu trnh ng mu. Bn cnh , cc protein c m ch yu l myosin phi hp vi actin to thnh actomyosin, chu trch nhim cho hot ng co c v.v.
III. M di truyn
Gene (DNA) c cu to t bn loi nucleotide, trong khi protein c cu to bi 20 loi amino acid. Vy vn t ra l, cc gene m ha cho cc sn phm protein ca chng bng cch no? Bng suy lun, ta c th suy on rng mi amino acid khng th c xc nh bi n v m gm mt, hai hoc bn nucleotide (v 41 = 4 hoc 42 =16 th cha m ha cho 20 amino acid, trong khi 44 = 256 th li

d tha qu nhiu) m c l phi l mt nhm gm ba nucleotide (43 = 64). Vi 64 kiu t hp b ba ho ra l tha m ho cho 20 loi amino acid. Nu nh th, mt amino acid c xc nh bi trung bnh ba b ba khc nhau. Vy phi chng m di tryn l m b ba? 1. Bng chng di truyn hc v m b ba Nm 1961, S.Brenner, F.Crick v L.Barnett phn tch chi tit nhiu th t bin ca phage T4 nhn c bng cch x l acridin, tc nhn gy cc t bin mt hoc thm mt cp base (chng 8), khng nh m di truyn l m b ba (triplet code) ng nh d on. n v m (coding unit) gm ba nucleotide xc nh mt amino acid nh vy c gi l codon. 2. Gii m di truyn Vic tip theo l xc nh xem mi amino acid c th c m ho bi mt hoc mt s b ba no. Cng trong nm 1961, M.Nirenberg v H. Matthaei ln u tin s dng mRNA nhn to c thnh phn base bit trc c tng hp bng enzyme polynucleotide phosphorylase (do Ochoa tm ra nm 1959) v h thng tng hp l dch chit t bo E. coli bao gm y cc yu t (cc ribosome, tRNA, amino acid, enzyme, ATP...) cn thit cho tin hnh gii m di truyn in vitro. Vi mRNA ch cha ton U, poly(U), chui polypeptide sinh ra ch cha ton phenylalanine (Phe). iu chng t UUU l b ba m ho ca Phe. Sau , Har Gobind Khorana tin hnh cc th nghim s dng cc mRNA tng hp c cha hai, ba hoc bn nucleotide c kt ni theo kiu lp li nh sau: (1) Vi mRNA nhn to cha hai base l poly(UC) hay UCUCUC..., s cha hai codon xen k UCU v CUC (ch rng s dch m in vitro khi u ti v tr ngu nhin). Kt qu l thu c mt polypeptide gm hai amino acid xp xen k nhau l serine v leucine, poly(Ser-Leu). (2) Vi mRNA tng hp gm cc b ba lp li s c dch thnh cc homopolypeptide. V d, poly(UUC) c th c c l (UUC-UUC), hoc (UCU-UCU), hoc (CUU-CUU) ty thuc vo v tr bt u dch m. V kt qu l c ba loi polypeptide c tng hp, poly(Phe) hoc poly(Ser) hoc poly(Leu). (3) Vi mRNA gm bn nucleotide lp li, v d poly(UAUC), th n c dch thnh polypeptide cha bn amino acid lp li l poly(Tyr-LeuSer-Ile). Tuy nhin, khi s dng cc poly(GAUA) v poly(GUAA) li khng cho kt qu. Qua so snh vi hng lot kt qu nhn c cho php xc nh c cc codon "v ngha" hay cn gi l cc tn hiu kt thc. T cc kt qu thu c nh vy Khorana xc nh c 'ngha' ca

phn ln codon c thnh phn base khng ng nht, v vic gii ton b h thng m di truyn (genetic code) c hon thnh vo thng 6/1966. Vi cng lao to ln Khorana v Nirenberg c trao gii thng Nobel nm 1968. T y cho php xy dng nn bng m di truyn (Bng 6.2) v rt ra c cc c tnh ca m nh c trnh by di y.
Bng 6.2 M di truyn (cho cc codon trn mRNA theo chiu 5'3')
3. Cc c tnh ca m di truyn - M di truyn l m b ba (triplet code), ngha l mt b ba nucleotide k tip m ho cho mt amino acid. Cc b ba trn gene v mRNA gi l codon (m) v b ba c trng ca tRNA c th khp vi codon ca mRNA theo nguyn tc b sung gi l anticodon (i m). - M di truyn khng gi ln nhau (non-overlapping): Mi codon l mt n v c lp, v thng tin ca mRNA c c theo mt chiu 5'3' (hnh 6.10) bt u t codon khi u.
3'
5'
Hnh 6.10 Cc phn t tRNA mang amino acid Ser (tri) v Tyr (phi) c m trn mRNA bng cch khp anticodon ca chng vi codon ca mRNA.

- M di truyn c tnh lin tc, khng b ngt qung (unpunctuated): Ngha l khng c khong h gia cc codon. - M di truyn c cc codon khi u (initiation) v kt thc (termination) nm gn hai u 5' v 3' ca mRNA ng vai tr l tn hiu khi u v kt thc tng hp chui polypeptide. Codon khi u AUG quy nh amino acid m u chui polypeptide l methionine ( vi khun l Nformyl-methionine). Cc codon kt thc (UAA,UAG hocUGA) khng xc nh amino acid no nn cn gi l cc codon v ngha (nonsense codon). - M di truyn c tnh n tr, r rng (unambigous): Mi codon xc nh mt amino acid duy nht, hoc xc nh s kt thc dch m. - M di truyn c tnh thoi ha (degenerate): C 61 codon c ngha (sense codon) trong khi ch c 20 loi amino acid; v vy mi amino acid (hay tn hiu kt thc) c th c xc nh bi nhiu hn mt codon. Cc codon cng xc nh mt amino acid nh th gi l cc codon ng ngha; chng thng khc nhau base cui, base 3' c gi l base thoi ha. V d, cc amino acid Arg, Ser v Leu mi ci c ti su codon ng ngha; Ala, Gly, Pro, Thr v Val mi ci c bn codon ng ngha (xem Bng 6.2). - M di truyn c tnh ph bin (universal), ngha l thng nht cho ton b sinh gii. 4. Nhng ngoi l so vi m di truyn "ph bin" Bn cnh tnh ph bin (universal) ca h thng m di truyn ni trn, cc nghin cu gn y cho thy mt vi chch hng m hu ht l xy ra trong cc b gene ty th (Bng 6.3). Tuy nhin, cc bo quan thc vt,
Bng 6.3 Cc ngoi l so vi m "ph bin" Ngun Ty th rui gim Ty th ng vt c v Ty th nm men
*(N = U, C, A hoc G)
Ty th thc vt bc cao Cc nhn Protozoa Mycoplasma
Codon UGA AGA & AGG AUA AGA & AGG AUA UGA CUN* AUA UGA UGA CGG UAA & UAG UGA
Ngha ph bin Kt thc Arginine Isoleucine Arginine Isoleucine Kt thc Leucine Isoleucine Kt thc Kt thc Arginine Kt thc Kt thc
Ngha mi Tryptophan Serine Methionine Kt thc Methionine Tryptophan Threonine Methionine Tryptophan Tryptophan Tryptophan Glutamine Tryptophan

s bin tp RNA (RNA editing) l ph bin v khng r rng, ch: Liu phi chng tt c cc trng hp sai lch so vi m ph bin thc vt l cc bin i tht hay l hu qu ca s bin tp RNA trc khi dch m. Mt vi thay i thnh thong cng xy ra cc b gene vi khun v b gene nhn ca cc eukaryote, nhng thng th lin quan vi cc codon kt thc. S phn b pht sinh chng loi ca cc thay i ny ch ra rng m di truyn vn cn ang tin ha. 5. S linh hot trong vic kt cp anticodon-codon Mc d c 61 codon c ngha nhng trn thc t trong mi t bo prokaryote v eukaryote ch c khong 45 phn t tRNA khc nhau. Tuy nhin, trong t bo ch c 20 loi amino acid c xc nh bi m di truyn, nn mt s loi tRNA phi mang cng mt loi amino acid. Cc bn sao tRNA nh th c th c trnh t anticodon ging nhau, trong trng hp chng c th thay th chc nng cho nhau. Cc tRNA khc th mang cc trnh t anticodon khc nhau v do vy nhn bit cc codon khc nhau; chng c gi l isoaccepting tRNA, v giu tng i ca chng c th nh hng ti cch thc s dng codon.
Bng 6.4 Cc nguyn tc kt cp linh hot anticodon-codon Base 5' ca anticodon G C A U I Base 3' ca codon C hoc U G U A hoc G U, C hoc A
Nm 1966, F.Crick a ra mt cch gii thch v s kt cp "lng lo" c th xy ra v tr th ba ca cc codon ng ngha. Theo Crick, hai base u tin ca mt codon phi c s kt cp chnh xc vi anticdon (theo nguyn tc b sung), cn base cui ca codon th c th "linh hot" (wobble), t c th hn so vi v tr bnh thng ca n hnh thnh nn s kt cp base bt thng vi anticodon. ngh ny c gi l gi thuyt linh hot (wobble hypothesis). c bit, Crick ngh rng mt base G trong anticodon c th cp khng ch vi C v tr th ba ca mt codon (v tr linh hot), m cn vi U. Hn na, Crick cn lu rng mt trong s cc nucleoside bt thng c mt trong tRNA l inosine (I), vn c cu trc tng t vi guanosine. Nucleoside ny thng thng c th kt cp nh G, v vy ta k vng n s cp vi C (kt cp base b sung) hoc U (kt cp base linh hot) v tr th ba ca mt codon. Nhng ng cn lu rng inosine vn cn c th c kiu kt cp linh hot khc, by gi ta bit l cp vi A v tr th ba ca codon. iu c ngha l,

mt anticodon c I v tr th nht v tim nng c th cp vi ba codon khc nhau c base cui l C, U hoc A. Cc th nghim sau ny khng nh iu d on ca Crick l hon ton ng v lit k cc kh nng kt cp base linh hot nh Bng 6.4. R rng l, hin tng kt cp linh hot ny lm gim ng k s lng cc tRNA cn thit dch m di truyn. V d, dch m cc codon (5'3') UUU v UUC m ho cho phenylalanine ch cn mt tRNAPhe mang anticodon (3'5') AAG.
IV. C ch phin m v sa i sau phin m

1. Cc RNA v c im chung ca phin m 1.1. S lc v cu trc cc RNA C bn loi ribonucleotide ni kt vi nhau bng cc lin kt phosphodiester to thnh cc chui polynucleotide ca RNA (v nguyn tc chung, c cp chng 5). Trong thnh phn base ca cc RNA, ngoi bn loi c bn adenine (A), uracil (U), guanine (G) v cytosine (C), cn pht hin mt s kiu base c sa i ph bin l trong cc tRNA (Hnh 6.11).
Dihydrouridine
Pseudouridine
1-methylguanosine
7-methylguanosine
1-methyladenosine
2-thiocytidine
5-methylcytidine
Ribothymine
Hnh 6.11 Cu trc mt ribonucleotide Uracil c trng ca RNA (tri) v mt s base sa i c th c trong thnh phn ca cc RNA.
C ba loi RNA c bn tham gia vo qu trnh sinh tng hp protein ca t bo cc sinh vt, l: RNA thng tin (messenger RNA = mRNA), RNA vn chuyn (transfer RNA = tRNA) v RNA ribosome (ribosomal RNA = rRNA). Ring cc rRNA, prokaryote c ba loi vi cc h s lng (sedimentation, c o bng n v Svedberg vi k hiu: S) l 5S, 16S v 23S; trong khi cc t bo eukaryote c bn loi: 5S, 5,8S, 18S v 28S. Ngoi ra, trong cc t bo eukaryote cn c cc RNA nhn kch thc ln v sai khc nhau gi l hnRNA (heterogenous nuclear RNA) vn l tin

thn ca cc mRNA, cc RNA nhn kch thc b snRNA (small nuclear RNA) c mt trong thnh phn ca cc enzyme splicing (xem phn sau), v cc RNA t bo cht kch thc b scRNA (small cytoplasmic RNA). Cu trc v chc nng ca cc RNA ny s c tho lun mc V. 1.2. c im chung ca phin m prokaryote v eukaryote Phin m (transcription) l qu trnh tng hp cc RNA khc nhau t thng tin di truyn cha ng trong DNA. Tr cc gene m ha protein trong cc operon vi khun, ni chung, cc RNA mi c tng hp ch l cc bn sao s cp (primary transcript) gi l cc pre-RNA. Cc pre-RNA ny phi tri qua mt qu trnh sa i tr thnh cc RNA trng thnh (mature) trc khi tham gia vo qu trnh sinh tng hp protein ca t bo. Qu trnh phin m DNA cc c im chung sau y (Hnh 6.12).
Hnh 6.12 S tng hp RNA trn mt si khun ca gene (DNA) di tc dng ca RNA polymerase.
(i) Din ra di tc dng ca cc enzyme RNA polymerase. (ii) Vng DNA cha gene c m xon cc b, v ch mt si n gi l si c ngha (sense strand) c dng lm khun (template) cho tng hp RNA. (iii) Phn ng tng hp RNA din ra theo nguyn tc b sung v c ko di theo chiu 5'3', ngc vi chiu ca si khun. (iv) Nguyn liu cho tng hp l bn loi ribonucleoside triphosphate: ATP, UTP, GTP v CTP. (v) Sn phm ca phin m l cc RNA si n (single RNAs). (vi) S khi u v kt thc phin m ph thuc vo cc tn hiu iu ho l cc trnh t DNA c th nm trc v sau gene c phin m. (vii) Qu trnh phin m c th chia lm ba bc, vn tt nh sau: M u (initiation) l s tng tc gia RNA polymerase vi vng promoter nhm xc nh si khun ca gene v tng hp vi nucleotide; Ko di (elongation) l giai an sinh trng tip tc ca chui RNA dc theo si khun cho n cui gene; v Kt thc phin m (termination) c trng bng s gii phng si RNA v RNA polymerase ra khi khun DNA.

2. Cc RNA polymerase ca prokaryote v eukaryote cc prokaryote, i din l E. coli, RNA polymerase hon chnh (holoenzyme) l mt phc hp gm nhn t sigma () v li enzyme. Nhn t sigma (sigma factor) gip cho RNA polymerase nhn bit v bm cht vo promoter c th bt u phin m ti v tr chnh xc, v li enzyme (core polymerase) ng vai tr chnh trong tng hp si RNA. cc eukaryote, c ba loi RNA polymerase I, II v III vi s phn b v chc nng chuyn ha khc nhau i vi b gene nhn, nh sau: RNA polymerase I trong hch nhn (nucleolus) phin m phc hp gene rRNA cho sn phm gm cc rRNA 18S, 28S v 5,8S; RNA polymerase II c trong dch nhn (nucleoplasm) phin m cc gene m ha protein cho sn phm l cc hnRNA/mRNA v c gene cho cc kiu snRNA (U1, U2, U4 v U5); RNA polymerase III c trong dch nhn phin m cc gene tRNA, rRNA 5S, v c snRNA U6. Ngoi ra, RNA polymerase ty th trong ty th v chu trch nhim tng hp tt c cc RNA ca ty th. 3. Cc promoter cc prokaryote v eukaryote
(a) Cu trc promoter cc Promoter structure in prokaryotes
5
prokaryote
mRNA
AUG
PuPuPuPuPuPuPuPu
[
-30 region TTGACA AACTGT -36 -31
-30
-10
+1
Promoter
]
-10 region TATAAT ATATTA -12 -7 Pribnow box 5 mRNA
transcription start site
+1
+20
T T G AC A
82 84 79 64 53 45%
TATA AT
79 95 44 59 51 96%
consensus t iu ha cc trnh sequences Cc nhn t phin m c th-trt DNA
(b) Cu trc promoter eukaryote Hnh 6.13 (a) Cu trc promoter cc prokaryote. (b) Cu trc promoter ca gene m ha protein trong nhn t bo eukaryote.

Cc vng khi ng (promoter) ni chung nm k trc gene v c cha cc on trnh t c th cho php RNA polymerase nhn bit v bm cht vo khi u phin m ti v tr chnh xc trn si khun ca gene. Vn ny tng i phc tp cc eukaryote, v vy y ta ch xt cc promoter ca cc gene m ha protein m khng cp cc loi promoter ca cc gene m ha cc RNA khc. Cc promoter ca cc gene m ha protein eukaryote v ca operon vi khun ni chung c cu trc kh tng ng nhau. on trnh t quan trng nht ca promoter c gi l hp TATA (TATA box) hay hp Pribnow (Pribnow box). i vi vi khun, l trnh t TATAAT (hoc tng t nh th) nm v tr "-10'' (Hnh 6.13); cn i vi cc gene m ha protein ca eukaryote, l trnh t TATAAA nm gn v tr "-30" v n c trng ring cho cc RNA polymerase II. (Cn lu l, tt c cc on tn hiu u c quy c trn si i khun ca gene, v chng c trnh t ging nh RNA c tng hp, ch khc l U thay cho T; cc k hiu '' v '+' ch cc v tr nm trc v sau v tr bt u phin m, hay cn gi l cc yu t upstream v downstream). Ngoi ra, trong cc promoter vi khun cn c trnh t TTGACA gn v tr ''-35'', gi l on nhn bit (recognition sequence). i vi cc gene m ha protein eukaryote, nm pha trc im bt u phin m chng 75 nucleotide c trnh t GGCCAAATCT, thng c gi l hp CCAAT (CCAAT box) - c l "hp cat"; n ng vai tr iu ha tc phin m. Ni chung, cc vng ny c bo tn cao v c th cho tng loi RNA polymerase nht nh, c gi l cc trnh t iu ha (consensus sequences). Cc t bin thay th base ti cc hp TATA, ngha l lm cho n bt ging vi trnh t c bo tn (v d, TATAAT TGTAAT) do s lm yu kh nng phin m ca promoter, gi l down mutation. Ngc li, cc t bin lm cho cc trnh t promoter tr nn ging vi cc trnh t iu ha (v d, TATCTT TATAAT), s lm mnh kh nng phin m ca promoter, gi l up mutation. 4. Cc giai on ca qu trnh phin m y ch cp mt m hnh n gin v qu trnh phin m E. coli. - Giai on bm v khi u: Sau khi RNA polymerase holoenzyme nhn bit v bm cht vo promoter, lm tho xon mt on chng 12 cp base ti y. Sau khi tng hp c mt vi nucleotide, nhn t sigma tch ra i vo mt chu k phin m khc, gi l chu k sigma (sigma cycle). - Giai on ko di: Enzyme li tin hnh ko di si RNA dc theo si khun. RNA polymerase li tin n u th DNA c m xon v phin m n y; v vng DNA c phin m ng xon tr li.

- Giai on kt thc: Khi qu trnh phin m tng hp xong hai on kt thc giu GC v AT nm ng sau gene th ti vng ui si RNA hnh thnh cu trc ''nt ci tc'' lm dng s phin m ca li RNA polymerase. Sau , di tc dng ca nhn t rho () c bn cht protein, si RNA va c tng hp v enzyme li c gii phng ra khi DNA khun. 5. S sa i sau phin m i vi cc mRNA eukaryote Nh cp, tr mRNA prokaryote ra, tt c cc RNA cn li d pro- hay eukaryote u phi tri qua qu trnh sa i sau phin m vi rt nhiu c ch tinh vi v phc tp khc nhau to ra cc RNA trng thnh tham gia vo qu trnh dch m; eukaryote, cc qu trnh ny xy ra trong nhn. c ci nhn h thng, y ta hy tm hiu mt t v cc c ch hon thin cc bn sao s cp mRNA (pre-mRNA) cc t bo eukaryote. 5.1. Gn thm "m" m7Gppp v "ui" poly(A) tr thnh phn t mRNA trng thnh trc khi i ra t bo cht lm khun cho dch m, tt c cc pre-mRNA ca cc gene m ha protein khc nhau t bo eukaryote, u c lp thm ci "chp" 7methylguanosinetriphosphate (m7Gppp cap) vo u 5' v "ui" poly(A) vo u 3' (Hnh 6.14). i vi cc gene m ha protein c vng m ha l lin tc (khng b gin on bi cc exon) nh cc gene histone chng hn, qu trnh hon thin mRNA dng li y. Loi ny chim khong 10%.
Trnh t m ho sau khi loi b cc intron
chp 5'm7Gppp
ui poly(A)
Hnh 6.14 Cu trc in hnh ca mt mRNA trng thnh eukaryote.
Cn lu rng, u 3' ca hu ht cc gene m ha protein eukaryote c cha trnh t AATAAA ng vai tr l tn hiu cho vic gn "ui" poly(A) vo u 3' ca mRNA. S phin m thng thng vn cn tip din sau khi i qua v tr polyadenyl ho ny. Trnh t tng ng vng cui 3' mRNA c phin m l AAUAAA bo hiu cho endonuclease nhn bit v ct chui RNA ti mt im xc nh nm sau n khong 10-30 base. Sau , mt enzyme khc l poly(A)-polymerase s lp thm vo u 3' ca mRNA mt dy adenine di khong 150-200 base gi l ui poly -A (poly [A] tail; Hnh 6.14). ui poly(A) c chc nng bo v mRNA khi b suy thoi v trong nhiu trng hp n cn kch thch s dch m. 5.2. S ct-ni i vi pre-mRNA ca cc gene phn an (split gene) V d, gene ovalbumin gm by intron xen k gia tm exon c di 7.700 cp base c E.Chambon phn tch trnh t nm 1981. Sau khi

enzyme splicing ct b cc intron v ni tt c cc exon trong mt qu trnh gi l x l RNA (RNA processing) th mRNA trng thnh c vng m ha protein di 1.872 base (Hnh 6.15).
Gene ovalbumin phin m Bn sao RNA s cp Chp 5'm7Gppp
1. lp chp 5' 2. lp ui poly(A)
ui poly(A)
loi b 5 intron (splicing) loi b 2 intron (splicing) xut ra t bo cht
mRNA trng thnh
Hnh 6.15 Phin m gene ovalbumin v s to thnh mRNA trng thnh.
Vic l gii c ch ct-ni (splicing) trong qu trnh x l pre-mRNA da ch yu trn hai s kin sau: (i) hai u mt ca mi intron c hai nucleotide rt n nh, gi l cc "trnh t chun", l 5'-GU......AG-3'. Mi intron ni chung c di khong 102-104 nucleotide; v (2) mt s snRNA cha trong thnh phn ca enzyme splicing cng c cc trnh t dinucleotide b sung vi cc trnh t chun trong intron. snRNA trong phc hp snRNP (small nuclear ribonucleoprotein) tng tc vi cc u mt ca mi intron, ko hai u mt xch li gn nhau to ra cu trc hnh vng, nh enzyme tin hnh ct b intron v ni cc exon li vi nhau.
mRNA tin thn ct pha 5'exon intron b ct b (thng lng)
ct pha 3'exon v splicing imct 5' im ct 3' ni cc u 5' v 3' ca cc exon mRNA trng thnh
Hnh 6.16 Mt m hnh v c ch ct-ni trong qu trnh x l pre-mRNA.
Vn t ra l s c mt cu cc intron trong cc gene phn on nh th c ngha g? Bi v vi li t chc nh vy lm cho trnh t m

ho ca gene b gin an, v trong qu trnh ch bin pre-mRNA ca n c th xy ra d l mt sai st nh cng to ra mRNA c khung c m b thay i. Ngi ta cho rng c khong 90% bn sao mRNA b suy thoi v ch li khong 10% mRNA trng thnh i ra t bo cht. Theo quan nim hin nay, s c mt ca cc intron trong cc gene phn on c th c cc vai tr sau: (i) cc intron nh l cc on m (spacer) to thun li cho s ti t hp gia cc exon bn trong gene; (ii) cc intron nh l cc vng m tch bit cc vng chc nng ca mt s protein; v (iii) cc intron nh l cc vng phn cch cho php cc exon c th c ct-ni c chn lc (alternative splicing) to ra cc mRNA trng thnh khc nhau v dch m thnh cc polypeptide khc nhau t mt gene; nh ni trc y, con ng ct-ni ny mang tnh c th cho tng m cc eukaryote bc cao.
V. Cu trc v chc nng ca cc loi RNA v ribosome

1. RNA thng tin (messenger RNA = mRNA) Cc mRNA l loi RNA quan trng nht dng lm khun cho tng hp polypeptide (Hnh 6.17); chng c cu trc 'm' vi kch thc khc nhau ty tng gene v u c ba phn chnh nh sau (xem thm Hnh 6.14):
5'-UTR vng m ha -'3UTR
(i) Vng dn u (5'UTR) tuy khng c dch m nhng cn thit cho s bm vo ca tiu n v ribosome b. (ii) Vng m ho (coding region) nm k sau 5'UTR, mang thng tin cu trc ca mt polypeptide, nu l mRNA ca eukaryote hoc
Hnh 6.17 Mt on ca phn t mRNA vi trnh t cc codon ca n.
mang thng tin ca nhiu polypeptide khc nhau (cch nhau bi cc on m khng c dch m), nu l mRNA ca prokaryote. (iii) Vng theo sau (3'-UTR) nm cui gene, khng c dch m. 2. RNA vn chuyn (transfer RNA = tRNA) Cc tRNA c chc nng chnh l mang amino acid v dch m cho mt codon trn mRNA (Hnh 6.10 v 6.18). Trong thnh phn ca chng thng c mt s base him tp trung cc vng thn (stem loop) nh: dihydro-uridine (DHU), inosine (I), ribothymidine (T) v pseudouridine

() (xem hnh 6.11). Cc tRNA u c cu trc khng gian ba chiu gn chc l do s kt cp cc base c th mt s on ca chng (Hnh 6.18). Ni chung, cc phn t tRNA gm khong 75-85 base, vi h s lng chng 4S; chng thng rt ging nhau nhiu on v sai khc ch yu l anticodon. Mi tRNA thng c 3-4 vng vi chc nng khc nhau: (i) vng DHU nhn bit aminoaxyl-tRNA synthetase; (ii) vng anticodon c m trn mRNA bng s kt cp anticodon-codon; (iii) vng "ph" (extra loop) c th khng c mt s tRNA; (iv) vng TC nhn bit ribosome i vo ng v tr tip nhn aminoaxyl-tRNA (v tr A); v (v) on mch thng CCA(3') l v tr nh ca amino acid.
V tr nh amino acid
Lin kt hydro gia cc cp base
Anticodon
Hnh 6.18 Cu trc bc ba (tri) v bc hai ca mt phn t tRNA.
3. RNA ribosome (ribosomal RNA = rRNA) Cc rRNA cng vi cc protein c th l nhng thnh phn cu trc nn cc ribosome -"nh my" tng hp protein ca t bo (Hnh 6.19). vi khun c 3 loi rRNA c cc h s lng l 23S, 16S v 5S, vi s lng nucleotide tng ng l 2904, 1542 v 120. t bo eukaryote c 4 loi rRNA vi cc h s lng l 28S, 18S, 5,8S v 5S. 4. Ribosome Tiu n v b
R 30S/40S R 50S/60S
ng knh
R 70S vi khun rRNA: 16S Protein: 21 phn t rRNA: 23S + 5S Protein: 34 phn t 18-20 nm
R 80S eukaryote rRNA: 18S Protein: >30 phn t rRNA: 28S +5S +5,8S Protein: >50 phn t 20-22 nm
Tv ln
Hnh 6.19 S cu trc mt ribosome vi hai tiu n v ln v b.
Mi ribosome c hai tiu n v b v ln (small and large subunits).

Tiu n v b c chc nng bm vo mRNA trc tin trong dch m. Tiu n v ln c cha peptidyl transferase xc tc hnh thnh cc lin kt peptide; n c cha hai v tr: v tr A l ni bm vo ca aminoacyltRNA v v tr P l ch dng tm ca peptidyl-tRNA. Hai tiu n v ny ch kt hp vi nhau to ra mt ribosome hot ng khi qu trnh dch m trn mRNA thc s bt u.
VI. C ch dch m (translation)

Dch m (translation) hay tng hp protein l mt qu trnh sinh hc quan trng din ra trong t bo cht, v ph thuc vo nhiu yu t; quan trng nht l mRNA, cc tRNA v ribosome. mRNA mang thng tin quy nh trnh t kt hp cc amino acid vo chui polypeptide, m vic dch m mRNA c thc hin bi cc aminoacyl-tRNA, cn ribosome ng vai tr n nh vic kt hp gia mRNA vi cc tRNA (Hnh 6.20). Qu trnh ny c chia lm hai giai on di y.
DICH MA
PROTEIN
Hnh 6.20 i cng v cc qu trnh phin m v sa i sau phin m din ra trong nhn, v dch m trong t bo cht t bo eukaryote.
1. Hot ho amino acid Qu trnh ny din ra trong bo tng v to ngun cc tRNA mang cc amino acid sn sng tham gia dch m. Mi amino acid c nh vo tRNA thch hp nh mt enzyme aminoacyl-tRNA synthetase c th. Trc tin, enzyme ny (E) xc tc cho phn ng ATP hot ho amino acid, vi s c mt ca Mg2+, to ra phc hp [Eaminoacyl~AMP]. RCH(NH2)COOH + ATP E*[RCH(NH2)CO~AMP] + PiP Tip theo, cng di tc dng ca enzyme , phc hp ny kt hp vi tRNA thch hp bng lin kt ng ho tr to ra aminoacyl-tRNA. E*[RCH(NH2)CO~AMP] + tRNA RCH(NH2)CO~tRNA + AMP 2. C ch ca qu trnh dch m (tng hp polypeptide) Bc 1: M u (initiation)

Qu trnh dch m bt u khi mt tiu n v ribosome b bm vo mRNA ti v tr ca codon khi u AUG. Lc ny mt phn t tRNA khi u c th mang methionine ( vi khun l formyl-Met) i vo v khp anticodon ca n vi codon m u ca mRNA. K , tiu n v ribosome ln bm vo tiu n v b to ra mt ribosome hot ng hon chnh. Lc ny Met-tRNA v tr P v v tr A trng; mt tRNA th hai (v d, tRNAVal) i vo v tr A v khp vi codon th hai (hnh 6.21A).
A
Lin kt peptid
D
Chui gm 8 amino acid
Tch v gii phng
Nhn t RF
Hnh 6.21 Qu trnh sinh tng hp chui polypeptide.
Bc 2: Ko di (elongation) Qu trnh ko di bt u sau khi lin kt peptide u tin c hnh thnh. Phn ng ny c xc tc bi enzyme peptidyl transferase, v kt qu l to ra mt peptidyl-tRNA v tr A (hnh 6.21B). Sau , ribosome lp tc chuyn dch sang mt codon mi dc theo mRNA theo chiu

5'3' (hnh 6.21C). Phn ng ny y phn t tRNA t do vn v tr P ra ngoi; lc ny peptidyl-tRNA nm v tr P v v tr A li trng. Mt chu k dch m mi li bt u, mt aminoacyl-tRNA th ba i vo v khp anticodon ca n vi codon ang trng v tr A, mt lin kt peptid th hai c hnh thnh, v ribosome li dch chuyn sang codon k tip. Qu trnh ni trn c din ra mt cch tun t dc theo mRNA lm cho chui polypeptide di dn ra cho n dch m xong codon 'c ngha' cui cng. Bc 3: Kt thc (termination)
Nhn t RF
Qu trnh tng hp chui polypeptide s dng li khi mt codon kt thc c a i din vi v tr A trng, vn c nhn bit bi mt protein kt thc gi l nhn t gii phng RF (release factor). S c mt ca n
Protein mi tng hp
Hnh 6.22 S kt thc dch m.
cng vi enzyme transferase ct ri chui polypeptide ra khi tRNA cui cng (hnh 6.21D) v phng thch hai tiu n v ribosome cng nh chui polypeptide v tRNA ra khi mRNA (hnh 6.22).
Hnh 6.23 So snh cc c ch tng hp v dch m mRNA cc t bo eukaryote (tri) v prokaryote.

Mt s im cn lu thm: (1) Trn y mi ch phn tch hot ng ca mt ribosome trn mRNA. Thc ra, trn mt mRNA c rt nhiu ribosome cng hot ng, gi l polyribosome hay polysome, to ra nhiu polypeptide ging nhau. (2) Trn nguyn tc, amino acid m u s c ct b khi chui polypeptide (sau khi tng hp c vi amino acid nh trong trng hp cc vi khun) hoc trc khi chui c tng hp y (nh trng hp eukaryote). Tuy nhin, cc eukaryote khng phi lc no amino acid m u ny cng b tch b, m trong mt s protein n vn c gi li. (3) Sau khi c tng hp, cc chui polypeptide s cp ny s c sa i v chuyn sang cc bc cu trc cao hn theo cch c th tr thnh cc protein hot ng chc nng (xem cc hnh 6.8 v 6.9). (4) Tham gia vo cc bc m u, ko di v kt thc cn c cc yu t protein, vi tn gi tng ng l cc nhn t m u (IF: initiation factor), nhn t ko di (EF: elongation factor), v nhn t gii phng (release factor) cng vi ATP, GTP v cc ion nh Mg2+, K+ v NH+4. (5) Trong cc t bo prokaryote, do khng c mng nhn v cc mRNA a cistron vn d khng phi qua sa i sau phin m, cho nn cc ribosome v cc aminoacyl-tRNA s bm vo u 5' ca mRNA bt u qu trnh dch m ngay trong khi u 3' ca n qu trnh phin m ang cn tip din. Ngc li, cc t bo eukaryote v c mng nhn phn cch v cc pre-mRNA cn phi tri qua cc cng on sa i phc tp sau phin m (tt c u din ra trong nhn), cn dch m din ra sau trong t bo cht (hnh 6.23). V th cho nn phin m v dch m r rng l hai qu trnh tch bit nhau c v c khng gian ln thi gian.
Cu hi v Bi tp
1. Ti sao ni gene l khi nim cn bn ca di truyn hc v tht kh c th a ra mt nh ngha chnh xc? Hy chng minh iu qua cc giai on pht trin ca di truyn hc cho n nay. 2. Bn hiu g v cc gene phn on (split gene)? Cho th d v s minh ho. Cc intron l g v c vai tr nh th no? Phn tch mt trng hp sa i (sau phin m) sn phm ca gene phn on lm sng t mnh mt gene c th xc nh nhiu hn mt loi polypeptide. 3. (a) Hy lm sng t nhn nh sau: Khc vi cc t bo prokaryote, s phin m v dch m cc t bo eukaryote l hai qu trnh tch bit c v khng gian ln thi gian. (b) Qu trnh tng hp v hon thin cc

RNA prokaryote v eukaryote ging v khc nhau nhng im no? 4. Trnh by vai tr chnh ca cc yu t tham gia vo qu trnh sinh tng hp protein ca t bo v phn tch c ch sinh tng hp protein da trn s tng tc gia mRNA, cc aminoacyl~tRNA v ribosome. 5. Bng suy lun, hy cho bit ti sao mt m di truyn ch c th c m ha theo nguyn tc b ba m khng th l kiu khc? Bng thc nghim iu c gii quyt nh th no? Tnh thoi ha ca m v nguyn tc kt cp linh hot c quan h vi nhau ra sao? Cho v d. 6. Phn tch s ph hp gia cu trc v chc nng ca cc loi RNA v mi quan h gia gene v protein. 7. Gi s tng hp c mt mRNA c thnh phn 75%U v 25%G. Khi s dng mRNA ny tng hp protein in vitro thu c cc amino acid trong cc protein vi cc tn s nh sau : Phe : Val : Leu : Cys : Gly : Trp = 1,00 : 0,44 : 0,33 : 0,33 : 0,15 : 0,11. Hy trnh by phng php v ch ra kt qu ca vic gii on cc codon cho mi amino acid ni trn (khng s dng bng m di truyn). Bit rng cc codon cng xc nh mt amino acid thng c hai nucleotide u ging nhau, v Cys c xc nh bi UGU. 8. (a) Hm lng GC ca DNA phage T3 l 53%. Bn s k vng hm lng G+C ca mRNA T3 ra sao? (b) Nu bit c hm lng purine ca DNA phage T3 v khng bit mch no lm khun, c th d on hm lng purine ca mRNA T3 hay khng? Ti sao, hoc ti sao khng? 9. Nu s dng cc phn t mRNA nhn to c thnh phn gm cc cm gm ba hoc bn nucleotide lp li di y tin hnh tng hp protein trong ng nghim, thnh phn amino acid thu c t cc polypeptide s nh th no? C trng hp no khng tng hp c protein hay khng? Ti sao? (a) (UUC)n ; (b) (UAC)n ; (c) (GAUA)n ; (d) (GUAA)n. 10. Nu s dng ba phn t mRNA tng hp c thnh phn gm cc base c kt hp ngu nhin theo t l sau y tin hnh tng hp protein in vitro, th thnh phn v t l cc amino acid c kt hp trong cc protein s nh th no? (a) 1U:5C (b) 1A:1C:4U (c) 1A:1C
Ti liu Tham kho

Ting Vit Hong Trng Phn. 1995. Mt s vn v Di truyn hc hin i (Ti liu BDTX cho gio vin THPT chu k 1993-1996). Trng HSP Hu.

Hong Trng Phn. 1997. Di truyn hc Phn t. NXB Gio Dc. Ting Anh Alberts B, Johnson A, Lewis J, Raff M, Robert K, Walter P. 2002. Molecular Biology of the Cell. Garland Science, NY. Cambridge Healthtech Institut. 2005. Gene Definition. www.healthtech.com Cooper DN. 2004. Gene structure, function and expression. http://www. cardiff.ac.uk/medicine/medical_genetics/study/medical_teaching/ McClean P. 1998. Definition of Gene. http://www.ag.ndsu.nodak.edu/plantsci/undrcour.htm Lewin B. 1999. Genes VI. Oxford University Press, Oxford. Lewis R. 2003. Human Genetics: Concepts and Applications. 5th ed, McGraw-Hill, Inc, NY. Palladino MA. 2002. Understanding the Human Genome Project. Benjamin/Cummings Publishing Company, Inc., Menlo Park, CA. Cambridge Healthtech Institut. 2005. RNA Glossary. www.healthtech.com Russell PJ. 2003. Essential Genetics. Benjamin/Cummings Publishing Company, Inc., Menlo Park, CA. Tamarin RH. 1999. Principles of Genetics. 6th ed., McGraw-Hill, Inc, NY. Twyman RM. 1998. Advanced Molecular Biology. BIOS Scientific Publishers Ltd/ Springer-Verlag Singapore Pte Ltd. Watson JD, Hopkins NH, Roberts JW, Steitz JA, Weiner AM. 1987. Molecular Biology of the Gene. 4th ed., Benjamin/Cummings Publishing Company, Inc, Menlo Park, CA. Weaver RF, Hedrick PW. 1997. Genetics. 3rd ed, McGraw-Hill Companies, Inc./ Wm.C.Browm Publishers, Dubuque, IA. Mt s trang web b sung Gene structure, function and expression. http://www.cardiff.ac.uk/medicine/medical_genetics http://www.oup-usa.org/isbn/019879276X.html

Chng 7
S iu ha Biu hin ca Gene

I. Cc nguyn l iu ha v mc kim sot phin m
Khng phi tt c cc gene u c biu hin lin tc. Mc biu hin ca gene khc nhau gia cc t bo hoc khc nhau theo giai on trong chu trnh t bo. Chng hn gene m ha cho hemoglobin c biu hin mc cao ch trong t bo tin th (precursor) ca t bo mu. Hot tnh ca gene khc nhau theo chc nng t bo. ng vt c xng sng nh chut, cha khong 200 loi t bo c phn ha chc nng khc nhau. Tt c cc t bo u cha cng thng tin di truyn, nhng t bo khc nhau ch nhng gene hot ng. Trong nhiu trng hp, hot tnh ca gene c iu ha mc phin m, c qua nhng tn hiu bt u bn trong t bo v c phn ng vi nhng iu kin bn ngoi. Tuy nhin thng tin di truyn c iu ha theo nhng cch khc nhau. Cc bc iu khin hot ng gene bao gm: - Cu trc li DNA, trong nhng thay i biu hin gene ph thuc vo v tr trnh t DNA trong genome. - iu ha phin m trong tng hp bn phin m RNA bng s iu khin s m u v s kt thc - Qu trnh ch bin RNA hoc iu ha qua qu trnh ct-ni trn RNA (RNA splicing) - iu ha dch m qu trnh tng hp chui polypeptid - S bn vng ca mRNA C s khc nhau ng k trong s iu ha biu hin ca cc gene eukaryote v prokaryote. Prokaryote l nhng c th n bo sng t do, sinh trng v phn chia trong iu kin thch hp v c cung cp y cht dinh dng. Do hot ng ca cc gene c iu ha do nhu cu ca t bo khi cn thit. Khc vi prokaryote, eukaryote l nhng c th a bo. Trong c th ang pht trin, mt t bo khng ch sinh trng v phn chia m t bo th h sau tri qua nhng thay i quan trng v hnh thi v ha sinh v duy tr trng thi bin i ca n. Hn na trong qu trnh pht trin phi, t bo eukaryote t chu nh hng ca mi trng hn so vi vi khun. Cui cng, c th trng thnh, s sinh trng v phn chia t bo ca hu ht cc loi t bo b ngng, mi t bo ch phi duy tr cc tnh cht ring bit ca n.

A. iu ha m tnh
im bm ca repressor
Phin m
Khng phin m Protein repressor bm vo
B. iu ha dng tnh
im bm ca nhn t hot ha
Khng phin m
Phin m Protein hot ha phin m bm vo
Hnh 7.1 M hnh iu ha m tnh (negative regulation) v iu ha dng tnh (positive regulation). A. Trong iu ha m tnh, protein c ch bm vo phn t DNA, ngn cn phin m B. Trong iu ha dng tnh, s bm vo ca protein hot ha phin m, kch thch phin m.
II. iu ha hot ng gene prokaryote

vi khun v phage, hot tnh ng m gene thng c iu khin qua phin m tng hp cc mRNA xy ra khi sn phm ca gene c cn n. C ch phn t cho mi m hnh iu ha hon ton khc nhau, nhng thng theo mt trong hai kiu chnh: iu ha m tnh v iu ha

dng tnh (Hnh 7.1). Trong h thng iu ha m tnh (Hnh 7.1A) mt protein c ch c mt trong t bo, ngn cn s phin m. Trong h thng cm ng c iu ha m tnh, protein c ch hot ng lm ngn cn phin m. Nhn t cm ng km hm cht c ch, cho php bt u phin m. Trong h thng c ch, protein aporepressor gn vi co-repressor to ra cht c ch c hot tnh, lm ngn cn phin m. Ngc li trong h thng iu ha dng tnh (Hnh 7.1B), s tng hp mRNA xy ra nu protein iu ha gn vo mt vng ca gene lm hot ha phin m. Nhng protein ny c xem l nhng nhn t hot ha phin m. iu ha m tnh v dng tnh khng loi tr ln nhau. Mt vi h thng l c iu ha m tnh v dng tnh, s dng c 2 h thng iu ha phn ng vi cc iu kin khc nhau trong t bo. iu ha m tnh l ph bin cho prokaryote, trong khi iu ha dng tnh li ph bin cho eukaryote. 1. Cu trc ca operon M hnh operon ca iu ha phin m C ch iu ha di truyn ca h thng lac (lac system) c gii thch bng m hnh operon ca Francois Jacob v Jacque Monod (1960) (Hnh 7.2). H thng s dng lactose gm 2 loi thnh phn: gene cu trc m ha protein cn thit cho s vn chuyn v chuyn ha lactose v cc yu t iu ha (gene c ch lacI, promotor lac P v operator lacO). Sn phm gene cu trc c m ha bi mt phn t mRNA a cistron (polycistronic). Gene Z m ha cho enzyme - galactosidase (thy phn ng lactose thnh galactose v glucose), gene Y m ha cho enzyme permease (cn cho vn chuyn lactose qua mng), gene A m ha cho enzyme transacetylase (vai tr chuyn ha lactose cha r). t bin promotor (lacP-) lm mt kh nng tng hp mRNA. Sn phm ca gene lacI l cht c ch, n bm vo trnh t cc base ca DNA cu to operator. Cht c ch bm vo operator, ngn cn s khi u phin m mRNA nh RNA polymerase. Cht cm ng (lactose) kch thch s sinh tng hp mRNA bng cch kt hp v lm bt hot cht c ch. S c mt ca cht cm ng lm cht c ch khng gn vo operator, promotor cho php khi u tng hp mRNA. Khi mi trng c lactose, lactose c chuyn vo t bo nh permease. Khi vo trong t bo mt s lactose (lin kt -1,4) c chuyn thnh allolactose (lin kt -1,6) nh -galactosidase. Allolactose l cht cm ng, n gn vo protein km hm, gy bin i cu hnh to

phc hp allolactose-repressor. Phc hp ny mt kh nng gn vo operator. Lc ny operon m ra, RNA polymerase bt u phin m t gene cu trc. Cc yu t iu ho Nhm gene cu trc [A]
Trc tip phin m
[B]
mARN Repress or protein
Repressor gn vo operator, ngn cn phin m
[C]
Phc hp cht cm ng-cht c ch khng th gn vo operator Inducer Khi khng lactose, RNA polymerase bt u phin m tng hp protein z, y v a
Lac mARN
Hnh 7.2 A Bn ca operon lac; B. S ca operon lac trng thi b km hm C. S ca operon lac trng thi c kch thch
galactosidase
Permeas e
Transacety lase
Khi mi trng khng c lactose, protein c ch c hot tnh gn vo operator, lm s phin m ca tt c cc gene cu trc ca operon lac b dng. S iu ha ca operon yu cu promotor nm chng ln mt phn hoc k st bn promotor ca gene cu trc, v n gn vi cht c ch

ngn cn phin m. 2. iu ha dng tnh operon lactose Chc nng ca -galactosidase trong chuyn ha lactose to ra glucose bng cch ct lactose (mt sn phm ct khc l galactose cng c th c chuyn thnh glucose nh enzyme ca operon galactose). Nu c glucose v lactose c mt trong mi trng sinh trng, khng cn hot ng ca operon lac. Trong mi trng c glucose, enzyme galactosidase khng c to thnh cho n khi glucose trong mi trng c s sng ht. S c mt ca glucose lm mRNA khng c tng hp, do khng c s tng hp -galactosidase, v s thm vo nhn t cm ng lm bt hot cht km hm. Mt yu t khc cn cho s bt u tng hp mRNA, hot tnh ca yu t ny c iu ha bi nng glucose. Glucose c nh hng c ch gin tip ln s biu hin ca operon lac. Nhng phn t nh adenosine monophosphate vng (cAMP) phn b rng ri trong m ng vt v trong cc c th eukaryote a bo, c vai tr quan trng lm cht trung gian hot ng hormone. Cht ny cng c trong t bo E. coli v nhiu t bo vi khun khc vi chc nng khc nhau. cAMP c tng hp bi enzyme adenyl cyclase, v nng ca cAMP c iu ha gin tip qua trao i cht glucose. Khi vi khun sinh trng mi trng cha glucose, hm lng cAMP rt thp. Trong mi trng cha glycerol hoc cc ngun carbon khng th i vo con ng ha sinh c s dng trao i cht glucose (con ng glycolytic) hoc khi vi khun b i ngun nng lng, nng cAMP cao. Hm lng glucose gip iu ha nng cAMP trong t bo v cAMP li iu ha hot tnh ca operon lac. E. coli cha protein nhn cAMP hay CRP (cyclic AMP receptor protein) cn c gi l protein hot ha d ha CAP (catabolite activator protein), c m ha bi gene crp. t bin gene crp v gene adenyl cyclase lm ngn cn s tng hp ca mRNA lac. iu ny cho thy chc nng ca CRP v cAMP cn thit cho tng hp mRNA Hnh 7.3 Cu trc ca cAMP lac. CRP v cAMP gn vo mt v tr khc to phc hp cAMP-CRP c biu hin. Phc hp ny l mt yu t iu ha hot ha h thng lac.

Tng hp ca mRNA l khng Repressor khng Phc hp cAMP-CRP
C Phin m khng
Hnh 7.4 Bn trng thi iu ha ca operon lac
Nhu cu v cAMP-CRP ph thuc vo h thng c ch lac, v t bin crp v adenyl cyclase khng th to mRNA lac ngay c khi c mt t bin gene iu ha (lacI-) v gene ch huy (lacO-). S d nh vy l v phc hp cAMP-CRP phi gn vo trnh t base trn DNA vng promotor xy ra phin m (Hnh 7.4). Khc vi cht c ch trong iu ha m tnh, phc hp cAMP-CRP l cht iu ha dng tnh. Cc th nghim iu kin in vitro cho thy: mRNA lac c tng hp ch khi c cAMP vng v khng c cht c ch. Khi vng mt phc hp cAMP-CRP, RNA polymerase ch bm lng lo vo promotor. V vy him khi dn n phin m v khng c s tng tc ng gia RNA polymerase v promotor. Nhng RNA polymerase c kch thch gn vo promotor khi cAMP-CRP c gn vo DNA. Nhng kt qu ny gii thch chc nng lactose v glucose cng nhau tham gia iu ha phin m operon lac nh th no. 3. iu ha m tnh operon tryptophan Operon tryptophan (trp) ca E.coli cha cc gene cu trc m ha cho cc enzyme tng hp amino acid tryptophan. Operon ny c iu ha theo cch sau: khi tryptophan c mt y trong mi trng sinh trng, s phin m ca operon b c ch. Khi s cung cp tryptophan b thiu, s phin m xy ra. S iu ha ca operon lactose tng t vi

operon lactose, v s tng hp mRNA c iu ha m tnh nh cht c ch. Tuy nhin, khc vi iu ha operon lac, tryptophan hot ng nh cht ng km hm, kch thch cht c ch gn vo operator ngng s tng hp. Operon tryptophan hot ng theo kiu c ch, iu ha m tnh. Tryptophan c tng hp qua cc giai on kh phc tp, mi giai on c s xc tc ca mt enzyme c bit. Cc gene m ha cho cc enzyme ny nm k nhau trn nhim sc th ca E.coli. l cc gene trpE, trpD, trpC, trpB, trpA. Cc enzyme c dch m t mt phn t mRNA a cistron. Vng m ha gene E c dch m trc tin. Pha trc trpE v u 5' c promotor, operator v 2 vng xp ln lt l leader (trpL) v on km hm phin m attenuator (trpa, khng phi l trpA). Gene c ch trpR nm xa operon, tng hp protein aporepressor, l cht km hm m ring n khng c hot tnh. Khi tryptophan d tha, n kt hp vi aporepressor to cht km hm c hot tnh, gn vo operator ca operon tryptophan lm dng phin m cc gene cu trc. Khi nng tryptophan thp, n tch khi phc hp km hm v aporepressor mt hot tnh. Lc ny operator m ra, RNA polymerase dch m 5 gene cu trc tng hp 5 enzyme tham gia tng hp ra tryptophan. (Hnh 7.5).
Phin m xy ra
trp P trp O trp L trp E trp D trp C
Aporepressor
Phin m Aporepressor khng bm vo operator
Phin m b c ch trp P
trp O
trp L
trp E
trp D
trp C
Khng phin m Phc hp Tryptophanaporepressor bm vo operator v c ch phin m Tryptophan Hot ha aporepressor
Hnh 7.5 iu ha ca trp operon E. coli A. Protein aporepressor khng bm c vo operator, phin m xy ra.

B. Khi c tryptophan, phc hp aporepressor v tryptophan lm cht c ch hot ng gn c vo operator, s phin m b km hm.
4. Phin m d (Attenuation) Kiu iu ha th hai c pht hin operon tryptophan c gi l attenuation. N dng s dch m iu khin s phin m. Khi c mt tryptophan ni bo, ngay c vi nng thp, s dch m mt phn vng leader ca mRNA ngay khi va c tng hp, kt qu lm dng s phin m trc khi gene cu trc u tin ca operon c sao chp.
[A ] [B ]
Kt thuc phin ma
Hnh 7.6 (A) S phin m ca leader trp; (B) Chi tit cu trc ca 2 codon trp vng 1-2
Phin m d (attenuation) l kt qu s tng tc gia cc trnh t DNA trong vng leader ca bn phin m trp. t bo kiu di, s phin m operon trp thng c bt u. Tuy nhin khi c mt mt lng nh tryptophan, hu ht phn t mRNA kt thc vng 28 base c bit trong trnh t leader. Kt qu ca s kt thc sm ny to phn t mRNA cha 140 nucleotide chm dt mt on ngn ca cc gene m ha cho cc enzyme trp. Vng 28 base xy ra s kt thc phin m sm nh th c gi l attenuator. Trnh t base ca vng ny thng c cc tnh cht im kt thc, gm dng on v vng (stem-loop) trn mRNA theo sau l trnh t ca 8 uridine. Trnh t leader c cc c im:

- Mt vng c codon AUG v pha sau l codon kt thc UGA, m ha cho mt polypeptide cha 14 amino acid c gi l leader polypeptide. - Hai codon tryptophan v tr 10 v 11 trn mRNA ca leader polypeptide. Trnh t lp li ngn ny c ngha trng iu ha. - Bn on ca RNA leader l vng 1, 2, 3 v 4 to thnh do kh nng kt cp ca cc base vi nhau. Cc base vng 1 kt cp vi vng 2, vng 3 kt cp vi vng 4 (Hnh 7.6). Khi s kt cp xy ra dng ny, s phin m kt thc on i qua uridine pha trc nucleotide 140. Kiu kt cp ny xy ra mRNA leader c tinh sch. - Mt kiu kt cp bin i c th xy ra, trong cc base vng 2 kt cp vi vng 3 nh cc cp base 2 vng ny gn nh b sung nhau (Hnh 7.6B). Qua m hnh kt cp base bin i ny (3-4 hoc 2-3), s t chc trnh t mRNA c th iu ha phin m qua dch m ca leader polypeptide (Hnh 7.7). Khi vng leader c phin m, s dch m leader polypeptid cng bt u. V c 2 codon ca tryptophan trong trnh t m ha, nn s dch m nhy cm vi s lng tRNAtrp a vo. Nu mi trng cung cp y tryptophan, ribosome trt qua codon tryptophan v i vo vng 2 (Hnh 7.7B). S c mt ca ribosome loi b kh nng kt cp ca vng khong 10 base mi pha ca codon ang dch m. S c mt ca ribosome vng 2 ngn cn n kt cp vi vng 3. Trong trng hp ny vng 3 kt cp vi vng 4, to ra im kt thc phin m. S phin m kt thc khi qua cc uridine nm pha sau vng 4. Khi s lng tRNAtrp khng , s dch m leader polypeptide b dng li t ngt cc codon tryptophan. S dng li ny ngn cn ribosome tin vo vng 2, v vy vng 2 c t do s kt cp vi vng 3 lm cn tr s hnh thnh cu trc kt thc. V vy phn t trp mRNA hon chnh c to thnh, cha c trnh t m ha cho gene cu trc. Tm li, attenuation l c ch iu ha tinh t trn c s iu ha m tnh: Khi tRNAtrp n cung cp cho s dch m leader polypeptide, s phin m b dng, cc trp enzyme khng c tng hp. Khi nng tRNAtrp qu thp, s phin m xy ra cho n ht, cc trp enzyme c to nn. Nhiu operon chu trch nhim tng hp cc amino acid khc (nh cc operon ca leucine, isoleucine, phenylalanine, histidine) cng c iu ha nh attenuator vi chc nng to ra vng kt cp bin i bn

phin m. operon histidine vng m ha ca leader polypeptide cha 7 codon histidine k nhau. operon phenylalanine vng m ha cho leader polypeptide cha 7 codon phenylalanin chia 3 nhm.
trp codons
Kt thc phin m
S kt cp trong RNA nng thp ca tryptophan
RNA polymerase
Phin m tip tc
S kt cp trong mRNA
S kt cp trong RNA nng cao ca tryptophan
Hnh 7.7 Phin m d (attenuation) ca operon trp E. coli A. mRNA t do c s kt cp base gia 1-2 v 3-4 B. nng cao ca tryptophan, ribosome tin n vng 2 v s kt cp 3-4 lm kt thc phin m C. nng tryptophan thp, ribosome vng codon trp cho php kt cp 2-3 v phin m khng b kt thc sau khi qua vng 4
iu ha kiu attenuation khng th xy ra eukaryote v eukaryote s phin m v dch m khng xy ra ng thi. S phin m xy ra trong nhn, cn s dch m xy ra t bo cht. iu ha operon lac v operon trp l v d v mt trong s cc c ch quan trng iu ha hot ng gene mc phin m ca prokaryote.
III. iu ha biu hin gene eukaryote

iu ha hot ng gene eukaryote phc tp hn nhiu so vi prokaryote. Lin quan n iu ha, gia prokaryote v eukaryote c mt s im khc bit: - eukaryote thng ch c mt chui polypeptide n c dch m t mt phn t mRNA hon chnh. mRNA a cistron (polycistronic) ch c prokaryote, khng c eukaryote. - DNA ca eukaryote gn vi protein histone to si chromatin, v gn vi protein phi histone. Ch c mt on nh DNA trn.

prokaryote, mt vi protein c mt trn nhim sc th b gp np, cn li hu ht DNA trn. - Mt on DNA quan trng ca eukaryote cha trnh t nucleotide lp li trung bnh hoc lp li cao. Mt vi trnh t lp li c xp ni tip thnh cc bn sao tip ni nhau, mt vi trnh t khc li khng xp ni tip. Vi khun cha on DNA lp li nh khc cc gene ca rRNA v tRNA nhn on v mt vi yu t di ng. Mt on ln DNA ca eukaryote khng c dch m. Hu ht trnh t nucleotide khng c m ha thnh protein. Cc eukaryote n bo, chng hn nm men l trng hp ngoi l i vi tnh cht ny. Mt vi gene eukaryote c biu hin v iu ha nh c ch cu trc li nhng on DNA nht nh theo mt phng thc c iu khin v tng s lng cc gene c bit ny khi cn thit. Cc gene eukaryote l gin on c phn thnh cc exon v intron. Cc intron c ct b trong qu trnh ch bin bn phin m RNA trc khi bt u dch m. eukaryote, mRNA c tng hp trong nhn v c chuyn qua mng nhn, vo t bo cht mi c s dng. T bo vi khun khng c nhn c tch bit vi t bo cht. 1. S bin i DNA Mt s gene ca eukaryote c iu ha bng s bin i DNA. Chng hn, nhng trnh t nht nh c th c khuych i hoc cu trc li trong genome hoc cc base c th b bin i v mt ha hc. Mt vi bin i c phc hi, nhng bin i khc li bn vng. Tuy nhin nhng thay i bn vng ny thng xy ra t bo sinh dng, v vy chng khng c truyn li cho th h sau qua dng giao t. 2. Cc promoter Tng t vi khun, cc promoter ca eukaryote cng nm pha trc im xut pht trn mRNA v c nhng trnh t c bo tn trong tin ha. Hp TATA nh hng cho mRNA bt u phin m nm pha trc im bt u phin m khong 30 bp ng vt c v v 60 n 120 bp nm men. Hp TATA hot ng c hiu qu cng vi 2 trnh t tng ng pha trc khong 40 bp l CCAAT v 110 bp l trnh t giu GC. S thay i hp TATA lm gim tc phin m. Hiu qu ca tc phin m c o bng s thay i ca tng base trong promoter. * S cu trc li DNA theo chng trnh (programmed DNA rearrangement) S cu trc li trnh t DNA trong genome l c ch bt thng

nhng quan trng, nh mt s gene c iu ha. 3. Nhng trnh t tng cng phin m (Enhancer) Cc th th ca hormone v nhng protein hot ha phin m khc gn vi trnh t DNA c bit c bit l enhancer. Trnh t enhancer kh ngn (thng ch 20 cp base) c tm thy cc v tr khc nhau quanh gene c iu ha. Hu ht cc enhancer nm pha di im bt u phin m (i khi cch xa nhiu kb). Nhng enhancer khc l cc intron nm trong vng m ha v mt vi enhancer thm ch nm u 3' ca gene. Enhancer l thnh phn nhy cm ca t chc gene eukaryote v chng cho php gene phin m ch khi no c nhn t hot ha phin m ng. Mt vi enhancer phn ng vi cc phn t bn ngoi t bo, chng hn hormone steroid to phc hp receptor-hormone. Nhng enhancer khc phn ng vi cc phn t c to thnh bn trong t bo (chng hn trong sut qu trnh pht trin). Nhng enhancer ny cho php cc gene di s iu khin ca n tham gia vo bit ha t bo (differentiation) hoc c biu hin theo cch c bit trong m. Nhiu gene di s kim sot ca cc enhancer khc nhau, v vy chng c th phn ng vi nhiu tn hiu phn t khc nhau c bn trong v bn ngoi.
Vng 5 Untranslated leader Introns Exons Vng 3
Enhancers
Promotor
Enhancers
Hnh 7.8 S t chc cc gene in hnh eukaryote bc cao
Qua s (Hnh 7.8) cho thy nhiu yu t di truyn c tm thy trong gene ca eukaryote in hnh. Phc hp phin m bm vo promotor bt u tng hp mRNA. Vng m ha ca gene (exon) b gin on bi mt hoc nhiu trnh t gin on (intron), cc trnh t ny s b loi b trong qu trnh ch bin RNA. S phin m c iu ha bi cc yu t enhancer, cc yu t ny phn ng vi cc phn t khc nhau c vai tr l tn hiu cm ng. Enhancer c mt c pha trn v pha di promoter. Mt vi enhancer c nhiu bn sao. Nhiu enhancer kch thch phin m bng cch hnh thnh vng DNA (DNA looping), lin quan n s tng tc gia cc vng cch xa nhau c lin quan dc si DNA. Cc nhn t cn thit cho phin m bao gm

protein hot ha phin m, n tng tc vi t nht mt yu t protein c trong mt hoc nhiu phc hp protein ln, nhiu yu t. Yu t protein trong phc hp ny c bit nh l yu t phin m chung (general transcription factors), v chng gn lin vi s phin m ca nhiu gene khc nhau. Nhn t phin m chung eukaryote c tnh bo tn cao trong tin ha. Mt trong s cc phc hp ny l TFIID, gm protein gn vi hp TATA (TATA-box-binding protein) TBP gn vi promotor trong vng TATA box. Ngoi TBP, phc hp TFIID cng gm mt s protein khc c gi l nhng nhn t gn lin vi TBP (TBP-associated factors) = TAFs. Chng hot ng nh cc cht trung gian qua nh hng ca nhn t hot ha phin m c truyn i. S phin m cng yu cu mt holoenzyme l RNA polymerase, cha pol III t hp t nht vi 9 yu t protein khc. nm men, nhng yu t ny cha nhn t phin m TFIIB, TFIIF v TFIIH cng nh nhng protein khc. 4. Trnh t bt hot gene (gene silencing) Trnh t bt hot gene c th nm trc hoc sau gene c iu ha khong vi trm cp base. Chng lm ngng qu trnh phin m bng cch bin i histone v DNA. 5. Promoter chn lc (alternative promoter) Mt vi gene eukaryote c hai hoc nhiu promoter hot ng trong nhng t bo khc nhau. Nhng promoter khc nhau ny dn n nhng bn phin m khc nhau cha cng vng m ha protein. V d Drosophila, gene m ha cho alcohol dehydrogenase, cu to ca n trong genome c ba vng m ha protein b gin on bi hai intron. S phin m u trng s dng mt promoter khc vi s phin m rui trng thnh. Bn phin m rui gim trng thnh c trnh t dn u 5' di hn. Nhng hu ht trnh t ny b loi b trong splicing. Promoter bin i lm s iu ha phin m c th khng ph thuc vo u trng hay rui gim trng thnh. 6. Splicing chn lc Cng promoter c s dng phin m mt gene, cc loi t bo khc nhau c th to sn phm c s lng khc nhau hoc to ra nhng protein khc nhau. iu ny l do cng mt bn phin m c th c qu trnh ch bin cc loi t bo l khc nhau. S tng hp -amylase chut, nhng phn t mRNA khc nhau c to ra t cng mt gene v nhng phn intron khc nhau b loi b trong qu trnh ch bin RNA. Tuyn nc bt ca chut to nhiu enzyme hn gan mc d cng mt trnh t m ha c phin m. mi

loi t bo, cng mt bn phin m s cp (primary transcript) c tng hp, nhng c hai qu trnh ch bin khc nhau (Hnh 7.9). Trnh t m ha bt u 50 bp bn trong exon 2 c to thnh nh ni vi exon 3 v nhng exon tip theo. tuyn nc bt bn phin m s cp c ch bin exon S ni vi exon 2 (exon L b loi i nh l intron 1 v 2). gan exon L ni vi exon 2, exon S b loi i cng vi intron 1. Exon S v exon L tr thnh nhng trnh t 5' bin i ca amylase mRNA v mRNA ny c dch m nhng t l khc nhau.
A. Bn phin m s cp Phin m amylase ca gan
Qu trnh ch bin t bo gan
mRNA amylase ca gan
B. Bn phin m s cp Phin m amylase ca tuyn nc bt Qu trnh ch bin t bo tuyn nc bt

mRNA amylase ca tuyn nc bt
Hnh 7.9 Sn phm cc phn t mRNA amylase khc nhau do qu trnh splicing khc nhau xy ra t bo tuyn nc bt v t bo gan ca chut.
Cu hi v Bi tp
1. S biu hin gene prokaryote v eukaryote c c im g? 2. Hy nu cc kiu iu ha ch yu. 3. Trnh by c ch hot ng ca operon lactose. 4. Operon l g? Nu ngha ca operon i vi s biu hin ca gene. 5. Gene m ha cho cht c ch (repressor) ca operon vi khun c cn thit phi gn gene cu trc khng? Ti sao? 6. So snh iu ha m tnh v iu ha dng tnh ca phin m. Cho v d. 7. Khi c mt glucose trong t bo E. coli, nng cAMP-CRP nh

th no? S gn ca cAMP-CRP vo DNA c nh hng n s gn ca repressor khng? 8. Nu ngha ca hin tng phin m d (attenuation). 9. Phn bit gia repressor v aporepressor v cho v d. 10. Th no l splicing chn lc? Cho v d minh ha.
Ti liu Tham kho

Ting Vit Phm Thnh H (2000). Di truyn hc. NXB Gio Dc. L nh Lng, Phan C Nhn (1998). C s Di truyn hc. NXB Gio Dc. Hong Trng Phn (1995). Di truyn hc Phn t. Trung tm o to T xa, i hc Hu. Ting Anh Anthony J. F. Griffiths, Susan R. Wessler, Richard C. Lewontin, William M. Gelbart, David T. Suzuki, Jeffrey H. Miller. 2004. An introduction to genetics analysis. W.H. Freeman Publishers. Harlt D.L., Jones E.W. (1998). Genetics - Principle and analysis. Jone and Bartlett Publshers. Toronto, Canada. Stansfield W.D. 1991. Schaums outline of theory and problems of genetics. McGraw-Hill, Inc., New York.

Chng 8
t bin gene, Ti t hp v cc Yu t Di truyn Vn ng

I. t bin gene
t bin gene l nhng bin i xy ra bn trong cu trc gene. Mi t bin gene dn n s thay i trnh t nucleotide to ra cc allele khc nhau. t bin gene c th xy ra do bin i ca trnh t nucleotide trong gene. t bin gene khng pht hin c khi quan st t bo hc. Trong t nhin, tt c cc gene u c t bin c gi l t bin t nhin hay ngu pht (spontaneous mutation). Cc t bin t nhin thng xut hin rt t. 1. Cc kiu t bin gene t bin gene hay t bin im: l cc bin i rt nh trn mt on DNA, thng lin quan n mt cp base n ca DNA hoc mt s t cp base k nhau. t bin im lm thay i gene kiu di (wild-type gene),. Thc t t bin im hu nh lm gim hoc lm mt chc nng ca gene hn l lm tng cng chc nng ca gene. V ngun gc, t bin im c phn ra lm t bin ngu pht (spontaneous) v t bin cm ng (induced). t bin cm ng: l dng t bin xut hin vi tn s t bin tng ln khi x l c mc ch bng tc nhn t bin hoc tc nhn mi trng c bit. t bin ngu nhin l t bin xut hin khi khng c s x l ca tc nhn t bin. t bin ngu nhin c tnh l t l c s ca t bin v c dng c chng ngun bin d di truyn t nhin trong qun th. Tn s t bin ngu nhin thp nm trong khong 10-5 10-8, v vy t bin cm ng l ngun t bin quan trng cho phn tch di truyn. Tc nhn t bin c s dng ph bin l ngun chiu x nng lng cao (high-energy radiation) hoc cc ha cht c bit. Cc dng t bin im: c hai dng t bin im chnh trong phn t DNA: + t bin thay th cp base (base substitution) + t bin thm-bt cp base (base insertion - base deletion) Cc t bin ny c th pht sinh do nh hng ca mi trng nh nh hng ca cc tc nhn gy t bin.

1.1. t bin thay th cp base Kiu t bin n gin nht l thay th mt base, trong mt cp nucleotide trong gene c thay th bng mt cp nucleotide khc. V d: A c thay th bng G trong si DNA. S thay th ny to ra s cp base G-T. ln sao chp tip theo to ra cp G-C trong mt phn t DNA con v cp A-T phn t DNA con kia. Tng t, t bin thay th A bng T trn mt si, to ra s kt cp tm thi T-T. Kt qu sao chp to ra T-A trn mt phn t DNA con v A-T trn phn t DNA con kia. Trong trng hp hp ny, cp base T-A l t bin v cp A-T khng t bin. Nu si gc DNA khng t bin c trnh t 5'-GAC-3', trn si t bin c trnh t 5'-GTC-3' v si kia khng t bin c trnh t 5'-GAC-3'. t bin thay th cp base c chia lm hai loi: + t bin ng hon (transition mutations): Nu mt t bin m base pyrimidine c thay th bng mt pyrimidine v mt purine thay bng mt purine. t bin ng hon c th l: T C hoc C T (Pyrimidine pyrimidine) A G hoc G A (purine purine) t bin o hon (Transversion): t bin lm thay mt pyrimidine thnh mt purine hay mt purine c thay th bng mt pyrimidine. Cc t bin o hon: T A, T G, C A hoc C G (Pyrimidine purine) A T, A C, G T hoc G C (Purine pyrimidine) Nh vy c th c 4 thay th kiu t bin ng hon v c n 8 thay th kiu t bin o hon. Nu cc thay th ny xy ra vi ngu nhin xc sut nh nhau, s c t l t bin: 1 ng hon : 2 o hon. Tuy nhin trong thc t, t bin thay th base c xu hng nghing v t bin ng hon, cho nn trong s cc t bin thay th base t pht th t l xy ra t bin l: 2 ng hon : 1 o hon 1.2. t bin thm hoc bt base (base-pair addition/deletion), cn gi l indel mutation (insertion-deletion). Trng hp n gin nht ca t bin

ny l thm hoc mt mt cp base n. i khi t bin lm thm hoc mt ng thi nhiu cp base. Hu qu ca t bin im n cu trc v s biu hin ca gene t bin im xut hin trong vng m ha chui polypeptide ca gene (a polypetide-coding part of a gene), chng hn t bin thay th base n c th gy nhiu hu qu, nhng tt c u c tc ng ln m di truyn theo 2 hng: lm thoi ha m di truyn hoc xut hin m kt thc qu trnh dch m. C cc dng: t bin ng ngha (synonymous mutations): t bin thay i mt codon m ha acid amine thnh codon mi m ha cho cng acid amin . t bin ng ngha cng c th xem l t bin im lng (silent mutations) t bin nhm ngha (missense mutations), i khi cn gi l t bin khng ng ngha (nonsynonymous mutations): codon m ha cho mt acid amin ny b thay i thnh codon m ha cho mt acid amin khc. t bin v ngha (nonsense mutations): codon m ha cho mt acid amin b thay i thnh codon kt thc dch m (translation termination/stop codon). Mc nh hng ca t bin nhm ngha v v ngha ln chui polypeptide khc nhau ty trng hp. Nu t bin nhm ngha thay th mt acid amin ny bng mt acid amin khc tng t v mt ha hc, c xem l t bin thay th bo th (conservative substitution). S thay i ny hu nh t nh hng n cu trc v chc nng protein. Ngc li, nu thay th bng mt acid amin khc v phng din ha hc gi l nonconservative substitution, hu ht u gy ra s thay i ln cu trc v chc nng protein. t bin v ngha s dn n s kt thc dch m sm. V vy chng gy ra hu qu tng ng trn chc nng protein. Nu t bin v ngha xy ra cng gn u 3' ca khung c m, kt qu t nh hng n protein. Tuy nhin nhiu t bin v ngha vng ny vn to ra cc sn phm hon ton b mt hot tnh.

Bng 8.1 t bin im mc phn t Kiu t bin mc DNA ng hon Kt qu v v d Purine c thay th bng mt purine khc, pyrimidine c thay th bng mt pyrimidine khc: AT GC, GC AT, CG TA, TA CG o hon Purrin c thay th bng mt pyrimidine hoc mt pyrimidine c thay th bng mt purine: AT CG, AT TA, GCTA, GCCG TAGC, TA AT, CG AT, CGGC Thm/Mt (Insertion-deletion) Thm vo hoc mt i mt hoc mt s cp base ca DNA (thm/mt base c gch di) AAGACTCCT AAGAGCTCCT AAGACTCCT AAACTCCT mc protein t bin ng ngha t bin nhm ngha Loi bo th Codon c bit m ho cho cng mt acid amin: AGG CGG Arg Arg Codon to thnh m ho cho amino acid khc M ho cho acid amin c cng bn cht ho hc: AAA AGA Lys Arg (kim) (kim) M ho cho amino acid khc v bn cht ho hc: UUU UCU Phenylalanin Serine K nc Phn cc Codon kt thc chui: CAG UAG Gln Stop
Loi khng bo th
t bin v ngha t bin dch khung
Thm vo mt cp base: AAG ACT CCT AAG AGC TCC T... Mt mt cp base: AAG ACT CCT AAA CTC CT...
Gene kiu di
t bin thay th base
t bin dch khung
Protein iu ha khng th bm
im t bin trong trnh t khng m ha
Hnh 8.1 Hu qu ca t bin im trong gene. Codon 1-4 nm trong vng m ha ca gene.
Ging vi t bin v ngha, t bin thm bt base gy hu qu trn trnh t polypetide k t im b t bin (Hnh 8.1). Trnh t trn mRNA c c theo tng khung gm ba base (codon) mt lc. Mt hoc thm base s lm thay i khung c trong qu trnh dch m t im b t bin cho n kt thc theo khung mi. V vy loi t bin ny c gi l t bin dch khung (frameshift mutations). t bin ny to ra trnh t acid amin k t im b t bin cho n kt thc khc vi trnh t acid amin gc. t bin dch khung gy ra s mt hon ton cu trc v chc nng ca protein bnh thng. Trng hp t bin xy ra trnh t iu ha v cc trnh t khng m ha khc (Hnh 8.1). Nhng phn ca gene khng trc tip m ha cho protein m cha nhiu im bm DNA ch yu cho protein xen vo, l nhng trnh t khng nhy cm cho s biu hin ca gene hoc cho hot tnh ca gene.

mc DNA, nhng im mt i (docking) gm nhng im m RNA polymerase v nhng nhn t gn kt ca n bm vo, cng nh nhng im m protein iu ha phin m c trng gn vo. mc RNA, nhng docking quan trng thm vo gm im bm ca ribosom (ribosome-binding site) trn mRNA vi khun, nhng im ni u 5' v 3' gn cc exon eukaryote v cc im c vai tr cho iu ha dch m v nh v mRNA n vng c bit trong t bo. Nhn chung hu qu chc nng ca bt k t bin im no vng nh th u ph thuc vo vic lm gin on (hoc to ra) mt im bm. t bin lm gin on nhng im c kh nng lm thay i phn biu hin ca gene da vo s thay i s lng sn phm c biu hin mt thi im nht nh hoc mt m nht nh. hay bng s thay i phn ng vi nhng tn hiu (cue) ca mi trng nht nh. Ngc li, t bin mt vi im bm c th hon ton ph hy mt giai on cn cho s biu hin bnh thng ca gene, nh im bm ca mRNA polymerase hoc l nhn t splicing. V vy n lm bt hot sn phm ca gene hoc ngn cn s hnh thnh sn phm. Cn phn bit gia nhng thay i xy ra ca mt t bin gene l s thay i trnh t DNA ca gene vi s thay i mc kiu hnh. Nhiu t bin im trong trinh t khng m ha lm t thay i hoc khng thay i trn kiu hnh nh t bin gia im bm DNA cho protein iu ha hoc thay i nhng im khc trong gene lm thay i chc nng ca chng. 2. Cc tc nhn gy t bin gene 2.1. Cc tc nhn gy t bin vt l - Phng x ion ha: cc tia phng x , , hoc tia X, cc chm tia neutron hoc proton lm bn cc in t gy ion ha v bin i DNA. - Phng x khng ion ha: tia t ngoi (UV) hoc nhit lm tng mc nng lng ca nguyn t. Tia t ngoi thng to dimer thymine gn hai thymine k nhau ca mt mch. 2.2. Cc tc nhn gy t bin ha hc - To sai hng khi sao chp: cc cht ng ng ca base (bromouracil, 2-amino purine) gn vo sai khi sao chp c chng. Cc cht mu acridine c th gn vo gia cc base ca mch xon kp lm mt hay thm vo mt base. - Tc ng trc tip ln gene khi khng c cc sao chp c th gy t bin im hay cc bin i A-T G-C hoc G-C A-T. 3. C ch phn t ca cc t bin gene

Khi kim tra dy t bin c gy to bi cc tc nhn t bin khc nhau cho thy mi tc nhn t bin c c trng bi mt c tnh t bin khc nhau hay "preference" v c mt dng t bin nht nh v mt im t bin nht nh, c gi l im d xy ra t bin (mutational hot spots). c tnh t bin nh th c ch ln u tin locus rII ca bacteriophage T4. Tc nhn t bin hot ng t nht qua ba c ch khc nhau: chng c th lm thay th mt base trong DNA; lm bin i mt base gy kt cp nhm vi mt base khc; lm sai hng mt base, dn n khng th kt cp vi bt k base no trong iu kin bnh thng. t bin thay th base: mt vi hp cht ha hc tng t nitrogen base bnh thng ca DNA, i khi chng c th gn vo DNA thay cho base bnh thng. Nhng cht nh th c gi l cc cht tng ng vi base (base analogs). Cc cht tng ng ny kt cp khng nh s kt cp ca cc base bnh thng. V vy chng c th gy ra t bin do gn vo mt nucleotide khng ng trong qu trnh sao chp.
Dng keto ph bin ca 5BU
Adenine
Dng keto ph bin ca 5BU
Dng enol him ca 5BU
Hnh 8.2 Chng minh mt vi kt cp nhm c th xy ra do kt qu ca s thay i mt tautomer thnh mt tautomer khc.
hiu hot ng ca cc cht tng ng base, trc ht cn phi xem xt khuynh hng t nhin ca cc base i vi s hnh thnh cc dng khc nhau. Mi base trong phn t DNA c th xut hin mt trong s nhiu dng c gi l tautomers, chng l cc ng phn khc nhau v tr v tr nguyn t v nhng lin kt gia cc nguyn t. Dng keto ca mi base thng c trong DNA (Hnh 8.2), trong khi dng imino v enol ca base l him. Tautomer imino hoc enol c th kt cp sai vi base to mt kt cp nhm (mispair). Kh nng kt cp nhm nh th gy ra t bin trong qu trnh sao chp c ch u tin bi Watson v Crick khi cc tc gi ny nghin cu cng thc v m hnh cu trc DNA.

S kt cp nhm c th sinh ra ngu nhin, nhng cng c th sinh ra khi base b ion ha. Tc nhn gy t bin 5-bromouracil (5-BrU) l cht tng ng vi thymine, c brome v tr carbon s 5 thay cho nhm CH3 ca thymine. Hot tnh ca n da trn qu tnh inolization v ionization. dng keto, 5-BrU kt cp vi adenin nh trng hp thymine. Tuy nhin, s c mt ca nguyn t bromine lm thay i mt cch c ngha s phn b electron vng base. V vy 5-BrU c th chuyn sang dng enol v dng ion, v n c th kt cp vi guanine nh trng hp cytosine to ra cp 5-BrU-G. Trong ln nhn i tip theo G kt cp vi C, to cp G-C thay cho cp A-T. Kt qu gy ra t bin ng hon. Tng t 5-BrU cng c th gy ra t bin ng hon A-T thay cho cp G-C. Mt ha cht gy t bin khc l 2-amino-purine (2-AP), l ha cht tng ng adenine, c th kt cp vi thymine. Khi b proton ha, 2-AP c th kt cp nhm vi cytosine, c th gy ra th h sau t bin ng hon G-C thay cho A-T do kt cp nhm vi cytosine trong ln sao chp tip theo. - Thay th base (base alteration) Mt vi tc nhn t bin khng gn vo DNA, m li lm bin i base gy ra s kt cp sai. Tc nhn alkyl c s dng ph bin nh l tc nhn t bin, chng hn nh ethylmethanesulfonate (EMS) v nitrosoguanidine (NG) gy t bin theo cch ny. Nhng tc nhn nh th s thm nhm alkyl (nhm ethyl trong trng hp EMS v nhm methyl trong trng hp NG) nhiu v tr trn c 4 base. Tuy nhin, t bin hu nh ch xy ra khi nhm alkyl c thm vo oxy s 6 ca guanine to ra O-6-alkylguanine. S alkyl ha ny dn n s kt cp nhm vi thymine (Hnh 8.3). Kt qu sinh ra t bin ng hon G-CA-T trong ln sao chp tip theo. Tc nhn xen vo gia (intercalating agents) l nhm tc nhn quan trong khc gy bin i DNA. Nhm ca cc hp cht ny bao gm proflavin, acridin cam v mt nhm cc hp cht ha hc khc. Cc tc nhn ny l nhm cc phn t bt chc cc cp base v c th xen vo gia cc base nit li chui xon kp DNA. v tr xen vo ny chng gy s thm vo hoc mt i mt cp nucleotide. - Sai hng base Mt s ln tc nhn t bin gy sai hng mt hoc nhiu base. V vy khng th kt cp vi base c trng. Kt qu lm cn tr s sao chp v DNA polymerase khng th tip tc qu trnh tng hp DNA qua

nhng base sai hng. E.coli qu trnh ny xy ra i hi hot tnh ca h thng SOS. H thng ny c kch thch nh l mt phn ng khn cp ngn cn s cht t bo khi DNA b sai hng nng.
Hnh 8.3 S kt cp nhm chuyn bit do t bin cm ng alkyl ho.
* C ch ca t bin ngu nhin t bin ngu nhin xy ra do nhiu nguyn nhn: gm sai hng trong qu trnh sao chp DNA, cc tn thng ngu nhin, s chen vo ca yu t di ng. t bin ngu nhin him nn kh xc nh c ch c bn. Tuy nhin, mt vi h thng chn lc cho php thu c t bin ngu nhin v phn tch mc phn t. T bn cht ca nhng thay i trnh t c th suy ra qu trnh dn n t bin ngu nhin. Nhng sai hng ngu nhin (spontaneous lesions) n DNA c th sinh ra t bin. Hai tn thng ngu nhin thng xut hin nht: depurine ho v deamin ho, trong depurine ho ph bin hn. Depurination do tc dng ca aflatoxin, lm mt mt base purine. Ngoi ra, qu trnh mt purine cng xy ra mt cch t nhin. Mt t bo ng vt mt ngu nhin khong 10.000 purine ca DNA trong mt th h t bo khong 20 gi 37oC. Nu tn thng ny c gi li, dn n sai hng di truyn ng k v trong qu trnh sao chp, v tr mt purine khng th nh r c loi base no. Trong nhng iu kin nht nh mt base c th chn vo to ra t bin.

Deamination ca cytosine to ra uracil. Uracil s kt cp vi adenin trong qu trnh sao chp, kt qu to ra t bin ng hon G-C A-T. Deamination 5-methylcytosine to ra thymine (Hnh 8.4). Qu trnh sao chp to ra t bin ng hon chuyn C thnh T.
Hnh 8.4 Deamination ca Cytosine (a) v 5-methylcytosine.
Ngoi 2 qu trnh gy sai hng nh trn, s oxy ha to ra cc base b sai hng l dng tn thng th ba.. Dng oxygen hot ng nh gc superoxid (O2-), hydrogen peroxide (H2O2) v gc hydroxyl (-OH) c to ra do sn phm ca qu trnh chuyn ha (aerobic metabolism). Cc dng ny c th gy tn thng oxy ha n DNA, kt qu to ra t bin. Cc sai hng trong sao chp DNA cng l ngun t bin khc. - Thay th base: sai hng trong sao chp DNA c th xy ra khi c mt cp nucleotide ghp khng chnh xc (nh A-C) to ra trong qu trnh tng hp DNA dn n s thay th mt base. - t bin thm vo v mt base: Mt loi sai hng sao chp khc dn n thm vo hoc mt mt hoc mt s cp base. Trong trng hp s base thm vo hoc mt i khng chia ht cho 3, s to ra t bin dch khung trong vng m ha protein.
II. Sa cha v bo v DNA

* C ch sa sai sinh hc T bo sng c hng lot h thng sai hng DNA theo nhiu cch

khc nhau. T l t bin t nhin thp do nh tnh hiu qu ca h thng sa sai ny. Sai hng ca h thng sa sai ny dn n t l t bin cao. Quang phc hot (photoreactivation) hay sa sai nh nh sng (light repair)
B khung DNA
nh sng trng Tia cc tm
Hnh 8.5 S to thnh v s loi b dimer thymine.
Sau khi x l tia t ngoi gy t bin, nu a ra nh sng th phn ln sai hng c phc hi nh enzyme photolyase. Enzyme ny gn vo photodimer ct n thnh cc monomer di tc dng ca nh sng mt tri c bc sng 320-370 nm. Sau phc hi cc base ban u. (Hnh 8.5). Sa sai bng lm mt nhm alkyl (dealkylation) Enzyme alkyltransferasecos th sa trc tip cc sai hng. Chng ct nhm alkyl t cht nitrosoguanine v ethylmethnesulfonate v gn vo v tr O-6 guanine. Enzyme methyltransferase ca E. coli c kh nng chuyn nhm methyl t cht O-6 methylguanine sang gc cistein trn phn t protein. Tuy nhin h thng sa sai ny c th bo ha nu mc alkyl ha cao. Sa sai bng ct b (excision repair pathway) Phn ln cc c ch sa sai khc thc hin theo li ct b (excistion repair) khng cn nh sng nh cc nuclease, sau thay vo cc base ng. C th xy ra theo nhiu cch:

- Ct cc base (base excision repair)
uvrABC exonuclease ct b on DNA 12 nucleotide
DNA polymerase I tng hp on DNA mi
Ligase ni ch h
Hnh 8.6 Sa sai bng ct b nucleotide.
S ct b cc base sai hng nh cc enzyme DNA glycosylase. Cc enzyme ny nhn bit cc base b bin i v cc im mt purine hay mt pyrimidine v thy gii lin kt N-glycosilic ni base vi ng. Ri enzyme AP endonuclease ct lin kt ng v phosphate gn base b bin i. Sau enzyme th ba, deoxyribophosphodiesterase loi b tng nucleotide k tip nhau on b hng. Sau , DNA polymerase lp y khong trng vi cc nucleotide b sung vi si khun cn li. Enzyme DNA ligase s gn cc khe h gia 2 u 3'-5' (Hnh 8.6).Trong t bo tn ti mt s DNA glycosylase. Chng hn, enzyme uracil-DNA glycosylase ct uracil khi DNA. Uracil to thnh do t bin mt nhm amin ngu nhin cytosine, dn n t bin ng hon thay C bng T. Enzyme ny pht hin ra uracil trn DNA nh l mt bt thng, chng s ct b v sa sai. - Ct cc nucleotide: S ct b vng c nhiu pyrimidine dimer c thc hin nh enzyme exonuclease (enzyme rch mch hay enzyme to khc trn DNA) nh phc hp 3 enzyme c m ha bi gene uvr ABC ca E. coli. Phc hp ny ct on 12 nucleotide trn mt mch: 8 nucleotide t mt u b sai hng v 4 nucleotide ca u cn li. Khong trng ca 12 nucleotide ny s c lp y nh enzyme DNA polymerase I da vo mch n b sung kia ca trnh t DNA gc. DNA ligase s gn vo cc khe h.

- c sa i vi cc base bt cp sai C ch c sa i vi cc base bt cp sai (proofreading for basepair matching) c thc hin trong sao chp DNA. Trong qu trnh sao chp, trc khi thc hin phn ng polymer ha ni cc nucleotide, cc nucleotide triphotphate mi phi bt cp b sung vi mch khun. Nu s bt cp sai xy ra, DNA polymerase s loi b nucleotide bt cp sai. Ngay c trc khi nucleotide mi rp vo, enzyme d li cp base cui, nu chng khng bt cp th s polymer ha tip theo b dng. Cp nucleotide u cui 3' bt cp sai s b loi b nh hot tnh exonuclease3'5' ca DNA polymerase. Khi bt cp ng, qu trnh polymer ha mi c tip tc. Hot tnh c sa i vi cc base bt cp sai l c tnh ca nhiu DNA polymerase m bo cho s ko di chnh xc ca mch ng c tng hp. + Sa sai da vo tnh tng ng (Homology-dependent repair system) Mt h thng sa sai quan trng pht hin tnh cht b sung i song song ca 2 mch n DNA phc hi on sai hng tr li trng thi bnh thng ban u. Trong h thng ny, on DNA sai hng b ct b v thay bng mt on nucleotide mi c tng hp b sung vi si khun i din. S sa sai xy ra qua si khun v nguyn tc ca sao chp DNA bo m s sa sai hon thnh vi chnh xc cao - l s gii phng sai hng (error-free). C 2 h thng ch yu loi b sai hng: H thng sa cha sai hng pht hin ra trc khi sao chp v h thng sa cha sai hng pht hin trong qu trnh din bin sao chp (sa sai sau sao chp). + Sa sai t mch kp (repair of double-strand break) Khi c 2 si ca chi xon kp b t cng mt v tr, c gi l t bin t mch i, c th gy ra sai hnh nhim sc th, lm cht t bo hoc to ra trng thi tin ung th. T bo s dng nhiu protein v con ng sa sai t gy mch i l thc hin ti t hp trong gim phn. Qu trnh sa cha do trao i cho trong gim phn xy ra nh sau (Hnh 8.7): Trn mt nhim sc th xy ra s t mch i v kt qu n mn cc u mt on ngn ca DNA si n. u 3' ca mt trong nhng si ny "xm ln" vo mt chromatid.

chromatid a chromatid A S b gy si i v ct u cui Chui DNA xon kp Chui DNA xon kp u 3
S b gy, xm ln v to vng S di chuyn vng v tng hp

Cu trc Holliday
Lp y khong trng v lin kt cc u t

Heteroduplex vi ch ghp nhm B gy v ni u 2 vi u 3
B gy v ni u 2 vi u 4
Sa cha A
Sa cha A
Kt qu khng trao i cho Trao i cho kp hon ton y s l mt y s l mt Nu khng sa cha octad 6:2 octad 6:2 A/a s to ra octad 5:3
Hnh 8.7 M hnh b gy si i nh trao i cho.
on xm ln lm mi cho tng hp cc base b mt ca n nh s dng si i song song ca chromatid nh l si khun. S tng hp mi ny s to ra mt vng si n lai vi mt si n khng xm ln. V vy to ra mt vng d hp t nh "Aa" v s dng nh mch khun khi phc cc base b mt trn si . DNA polymerase s lp y ch trng v enzyme ligase s ni cc u mt xy ra trong cu trc c bit ging vi trao i cho 2 si n. Cu trc ny cng cha cc on bt cp khng tng ng n gin. Trao i cho si n c gi l cu trc Holliday (Holliday structure) do Holliday pht hin vo nhng nm 1960. H thng SOS

t bo vi khun hoc t bo eukaryote b sai hng nng do chiu tia uv, tia X hoc do tc dng ca cc ha cht gy t bin, h thng sa sai khn cp c khi ng. E. coli, h thng ny c lin quan vi hai protein c m ha bi gene lexA v recA. Protein lexA l mt cht c ch, n gn vo hp SOS, chng lp cc promotor ca cc gene SOS, ngn cn s phin m nhm cc gene ca h thng SOS. Mt vi sn phm ca DNA b tn thng s lm hot ha enzyme protease recA. Protein recA b hot ha s ct b protein lexA, cho php cc gene ca h thng SOS phin m. Phn ng ca h thng SOS xy ra trong thi gian ngn nhng phc tp. N bao gm cc qu trnh lm tng hot tnh ti t hp, thay i trong khi s sao chp, c ch nuclease v kch thch phc hi sao chp v chuyn sai hng thnh sa sai p sp (error-prone replication). T bo by gi s xy ra s sao chp DNA nhanh hn bnh thng. Nu sa sai khng kp, t bo phi chp nhn hoc b t bin hoc b cht. III. Cc yu t di truyn vn ng (Transposable genetic elements) 1. Cc yu t di truyn vn ng prokaryote Trnh t on xen (insertion sequence) ca vi khun Trnh t xen on l mt on DNA ca vi khun di chuyn t mt v tr trn nhim sc th n v tr mi trn cng nhim sc th hoc trn nhim sc th khc. Khi xen vo gia gene, yu t IS lm gin on trnh t m ha v lm bt hot s biu hin ca gene. Mt s trng hp, c tn hiu kt thc phin m v dch m, yu t IS lm cn tr s biu hin sau promotor trong cng operon.. Yu t IS c tm thy u tin operon gal ca E. coli, chia lm bn nhm: IS1, IS2, IS3 v IS4. Chng c th phn b ri rc trn nhim sc th chnh ca vi khun v trn cc plasmid. V d yu t IS1 c khong 5-8 bn sao trn nhim sc th vi chiu di 768 bp. Tt c cc yu t IS u cha on DNA m ha cho protein, c gi l transposase, l enzyme cn thit cho s di chuyn ca yu t IS t mt v tr trn nhim sc th n v tr khc. on gene ny nm gia 2 on lp li o ngc (inverted repeat - IR) ngn. V d yu t IS1 c khong 5-8 bn sao trn nhim sc th vi chiu di 768 bp, on lp li o ngc c kch thc 18-23 bp, yu t IS2 c 5 bn sao v cc yu t IS khc. Yu t IS l nhng vng ca cc trnh t xc nh, chng l nhng v tr xy ra trao i cho. V d: S ti t hp ca plasmid v nhim sc th ca E. coli to ra nhng chng Hfr xy ra qua trao i cho n gia yu t IS trn plasmid v yu t IS trn nhim sc th.

Bng 8.2. Mt vi trnh t xen vo v kch thc ca chng Trnh t xen vo IS1 IS2 IS3 IS4 IS5 S bn sao trong E. coli 5-8 bn sao trn nhim sc th 5 bn sao trn nhim sc th, 1 bn sao trn plasmid 5 bn sao trn nhim sc th, 2 bn sao trn plasmid 1-2 bn sao trn nhim sc th, Cha bit Chiu di (bp) 768 1327 1400 1400 1250 on lp li o ngc (bp) 18-23 32-41 32-38 16-18 Ngn
1.1. Gene nhy ca prokaryote Cc yu t IS ring l khng ch c kh nng t di chuyn m khi hai yu t ny nm gn nhau th chng c th vn ng nh mt n v hon chnh v mang theo cc gene nm gia chng. Cu trc phc tp ny c gi l transposon. C hai kiu transposon vi khun:
(a) Transposon hn hp
(b) Transposon n gin
Hnh 8.8 c trng v cu trc ca transposon hn hp (composite transposon) v transposon n gin (simple transposon).
Transposon hn hp (composite transposon) cha nhiu gene nm gia 2 trnh t IS gn nhau, c hng ngc nhau to ra trnh t lp li o ngc (inverted repeat = IR). Mt trong 2 yu t IS m ha cho transposase xc tc cho s chuyn v ca c transposon. Chng hn Tn10 l transposon hn hp mang gene m ha cho tnh khng khng sinh tetracyline. Gene ny nm gia hai yu t IS10 c hng ngc nhau.

Transposon n gin (simple transposon) gia cc trnh t IR, nhng nhng trnh t ny ngn (<50 bp) v khng m ha cho transposase. S chuyn v ca chng khng phi l kt qu ca s lin kt vi yu t IS. Cc transposon n gin m ha transposase ring thm vo mang cc gene ca vi khun. Tn3 l mt transposon n gin (Hnh 8.8). Transposon hn hp v transposon n gin u cha cc gene thm vo lin quan n chc nng mi t bo vi khun. C hai loi ny thng c gi chung l transposon. Transposon di hn yu t IS, thng cha vi kb), chng cha cc gene m ha cho protein thm vo. 1.2. C ch ca s chuyn v u tin, transposase ct vt hnh ch chi qua 5 cp base (khc vi s ct ca enzyme restriction endonuclease) vi tr DNA mc tiu (target site DNA) (Hnh 8.9). Tip theo l s hi nhp ca transposon qua trung gian ca transposase, transposon xen vo gia cc u mt ca ch chi. u li ra ca si n c s dng nh l khun tng hp si b sung th hai. S gn vo to s sao chp 5 cp base, c gi l s sao chp im mc tiu (target site duplication). Hu ht cc yu t di ng ca prokaryote u s dng mt trong 2 c ch chuyn v l sao chp (replicative) v bo th (conservative) hay khng sao chp. Trong con ng sao chp (nh Tn3), mt bn sao mi ca yu t di ng to ra khi chuyn v, kt qu l mt bn sao v tr mi v bn sao cn li v tr c. Trong con ng bo th (nh trng hp Tn10) khng c s sao chp. Thay vo , yu t c ct ra t nhim sc th hoc plasmid v c gn vo v tr mi. Con ng ny cn c gi l con ng "ct v dn" (cut and paste) 2. Cc yu t di truyn vn ng eukaryote Cc yu t di truyn vn ng eukaryote chia lm 2 nhm: nhm l cc retrotransposon v nhm 2 l cc DNA transposon. 2.1. Cc retrotransposon Gery Fink v cs. l nhng ngi u tin s dng nm men nghin cu iu ho hot tnh gene eukaryote. Cc tc gi ny phn lp c hng ngn t bin gene HIS4 m ha enzyme tham gia tng hp histidine. Trong s hn 1.500 t bin ngu nhin HIS4 c tm thy c 2 t bin c kiu hnh khng bn vng. Cc t bin khng bn vng ny c tn s phc hi li dng kiu di cao 1.000 ln hn cc t bin HIS4 khc. Nhng t bin ny cho on DNA ln xen vo gene HIS4, s xen vo ny c thc hin do mt trong cc yu t l Ty ca nm men. C 35

bn sao ca yu t xen on gi l Ty1 genome ca nm men.
Transposase ct on DNA mc tiu
Yu t transposon xen vo
Lm y cc ch trng
Yu t lp li trc tip 5 cp base 2 u Hnh 8.9 S nhn i on trnh t DNA ngn im xen vo.
Vic to dng nhng yu t ny t cc allele t bin cho thy xen on ny khng ging vi yu t IS hoc transposon ca vi khun. Thay vo chng c c tnh ca retrovirus (virus ca ng vt). C s ging nhau trong cu trc v thnh phn gene ca retrovirus v yu t Ty1 c phn lp t t bin HIS4. Ging vi retrovirus, transposon ca nm men c lp on cui di (long terminal repeat sequence) LTRs, cha hng trm cp base, c gi l trnh t nm 2 pha on m ha, c hai u cha gene gag v gene pol. Retrovirus c t nht 3 gene m ha cho 3 protein trong qu trnh sao chp: gene gag m ha cho mt protein c vai tr lm bin tnh RNA genome. Gene pol m ha enzyme reverse transcriptase. Gene env m ha cho protein v. Yu t Ty ch cha gene gag v gene pol, khng cha gene env. (Hnh 8.10).

M hnh v s chuyn v nh retrotransposon. Mt bn phin m RNA t retrotransposon di tc dng ca enzyme phin m ngc to thnh DNA nh enzyme reverse transcriptase c m ha bi retrotransposon. Bn sao DNA c chn vo v tr mi trn b gene.
on lp li trc tip 5 bp ca DNA mc tiu Phin m NST khc
Bn sao Ty1 xen vo
Dch m Phin m ngc Enzyme phin m ngc
Hnh 8.10 S chuyn v nh retrotransposition.
Vo nm 1985, J.Bocke v G. Fink chng minh, yu t Ty1, ging vi retrovirus, thc hin vic di chuyn qua trung gian RNA. Chng bt u bng bin i yu t Ty1 ca nm men c to dng trn plasmid. Trc tin mt u mt ca yu t, c s xen vo mt promotor c hot ha nh thm galactose vo mi trng. Th hai, mt intron t mt gene khc ca nm men c a vo vng m ha ca transposon Ty. S thm vo galactose lm tng tn s chuyn v ca yu t Ty bi bin i. iu ny lm tng s lng RNA, v galactose kch thch phin m RNA Ty bt u t promotor nhy cm galactose. Nhiu t bin ngu nhin c phn lp rui dm cho thy cng cha retrotransposon. Yu t copia ca rui dm cu trc tng t vi yu t Ty ca nm men. Chng xut hin t 10-100 v tr trn b gene ca rui dm. Nhng t bin nht nh ca rui dm l kt qu ca s xen vo ca yu t copia v nhng yu t khc. Chng hn, t bin white-apricot (wa) v mu mt ca rui dm to ra do s xen vo yu t ca h copia locus white.

S xen retrotransposon LTR vo cc gene thc vt nh ng, cng gp phn to ra cc t bin ngu nhin. 2.2. DNA transposon Nhiu yu t di ng c tm thy eukaryote chuyn v bng c ch ging vi vi khun. Bn thn yu t IS v transposon hoc l bn sao ca chng c th xen vo v tr mi trn genome. Cc yu t di chuyn theo cch ny c gi l yu t nhm 2 hoc DNA transposon. Yu t di ng c McClintock pht hin u tin ng l cc DNA transposon. Tuy nhin tnh c trng phn t ca DNA transposon u tin li l yu t p rui dm. Yu t p c pht hin khi nghin cu th lai mt kh nng sinh sn (dysgenesis), hin tng xy ra khi lai rui ci thuc chng phng th nghim vi rui c qun th t nhin. Trong php lai nh th, chng phng th nghim c xem l c kiu t bo M (M cytotype) v ging gc t nhin c xem l c kiu t bo P (P cytotype). Php lai ca M (ci) x P (c) cho th h sau kiu hnh dng mm vi gm dng bt th vi t l t bin, tn s sai hnh nhim sc th v khng chia ly nhim sc th cao. Php lai nghch khng xy ra hin tng thoi ha ging (dysgenics). t bin dysgenics khng bn, chng c th phc hi dng kiu di hoc bin i thnh allele t bin khc vi tn s cao. S ging nhau v t bin khng bn vng rui dm v ng a n gi thuyt t bin dysgenic c gy ra do s xen yu t di ng vo nhng gene c bit, lm chng b bt hot. Chng hn hu ht cc t bin trn rui dm do xen yu t di ng vo gene white. Nhng yu t nh vy gi l yu t p, c t 30-50 bn sao trn genome chng P v hon ton vng mt chng M. Yu t P c chiu di t 0,5-2,9 kb. Kch thc khc nhau ny do do nhiu yu t P khng y m b mt on gia. Yu t P vi kch thc y tng ng vi transposon n gin vi khun, c u mt l on lp o ngc ngn (31 bp) v n m ha cho transposase. Gene m ha cho transposase eukaryote cha 3 intron v 4 exon. ng, yu t Ac v Ds c th di chuyn trn nhim sc th , tc ng ln cc gene kim tra mu sc ca cc ht aleuron. Cc yu t ny c phn lp cho thy c lin quan vi DNA transposon ca vi khun v ca cc eukaryote khc. Ging vi yu t P ca rui dm, yu t Ac c on cui lp li o ngc m ha cho transposase, yu t Ds li khng m ha cho transposase. Khi Ac trn genome, transposase ca n c th bm vo u mt ca c yu t Ac v Ds v khi ng s chuyn v ca chng..

Yu t Ac v Ds l thnh vin ca h transposon n gin. Ngoi ra, cn c cc h yu t di ng khc ng. Mi h cha yu t t ng m ha cho transposase c th chuyn c yu t trong cng mt h, nhng khng th chuyn n cc h khc v transposase ch c th bm vo u mt ca cc thnh vin trong h. ngi, yu t di ng chim mt na genome ngi. a s yu t di ng thuc 2 dng retrotransposon l yu t nhn ri rc kch thc di (long interspersed nuclear element) LINEs v yu t nhn ri rc kch thc ngn (short interspersed nuclear element) SINEs. LINE di chuyn nh retrotransposition s dng yu t m ha reverse transcriptase, nhng li thiu mt vi tnh cht v cu trc ca yu t ging retrovirus bao gm LTR. SINE c m t nh l LINE khng t ng, v chng c tnh cht cu trc c trng ca LINE nhng khng m ha cho reverse transcriptase ring. Ngi ta cho rng chng di chuyn c nh enzyme reverse transcriptase c m ha bi LINE trong genome. SINE ngi c gi l trnh t Alu v n cha trnh t im ct ca enzyme ct hn ch Alu. Genome ca ngi cha hn 1 triu trnh t Alu ch mt phn hoc ton b, chim hn 10% genome ca ngi. Trnh t Alu y c kch thc 200 nucleotide. DNA genome mang yu t di ng ln hn 20 ln DNA m ha cho tt c protein ngi. Tn s t bin ngu nhin do xen vo yu t nhm 2 l thp cha n 0,2% trong tng cc t bin ngu nhin. Trong khi nhng t bo ng vt khc nh chut do xen retrotransposon ln n 10% t bin ngu nhin, cao hn 50 ln ngi c l lin quan vi hot tnh ca yu t retrotransposon cao hn chut.
Cu hi v Bi tp
1. V sao phn ln cc t bin nh hng n cc gene cu trc thng l ln so vi cc allele hoang di? 2. S hnh thnh t bin dch khung din ra nh th no? 3. Cc ha cht gy t bin c c im g? 4. Gii thch c s t bin ca cc tc nhn gy t bin sau: 5bromuracil, acid nitr v acridin. Xc nh xem chng to ra dng t bin no (ng hon, o hon hay dch khung)? 5. Hy m t mt loi sai hng ngu nhin dn n t bin. 6. Phn tch s ging nhau v khc nhau gia cc kiu transposition.

7. Cc trnh t o ngc c vai tr g trong transposition? 8. Cho mt chui trnh t nucleotide trn mRNA nh sau: Dng hoang di: ... 5' AAUCCUUACGGA 3' ... Dng t bin: .... 5' AAUCCUACGGA 3' ... Hy cho bit: Sai hng trn xy ra do loi t bin no? 9. t bin xy ra trong trnh t nucleotide do kt cp nhm nh sau: 5' AGCTGCCTT 3' 3' ACGATGGAA 5' (mch khun) mRNA Hy cho bit: Amino acid no s c tm thy codon c nucleotide b thay i nh trn? 10. bp, tn s t bin ca locus R (mu cy) rt cao: 492 t bin trn 106 giao t. Gene to mu ca ni nh Pr c tn s t bin l 11 t bin trn 106 giao t. Hy cho bit: Cn phi phn tch bao nhiu cy mi tm c 1 t bin kp ca 2 gen trn?
Ti liu Tham kho

Ting Vit Phm Thnh H. 2000. Di truyn hc. NXB Gio Dc. L nh Lng, Phan C Nhn (1998). C s di truyn hc. NXB Gio dc. Hong Trng Phn. 1995. Di truyn hc phn t. Trung tm o to T xa, i hc Hu Ting Anh Anthony J. F. Griffiths, Susan R. Wessler, Richard C. Lewontin, William M. Gelbart, David T. Suzuki, Jeffrey H. Miller. 2004. An introduction to genetics analysis. W.H. Freeman Publishers. Harlt D.L., Jones E.W. 1998. Genetics - Principle and analysis. Jone and Bartlett Publshers, Toronto, Canada. Stansfield W.D. 1991. Schaums outline of theory and problems of genetics. McGraw-Hill, Companies, Inc., United States of America. Watson D.J, Baker T.A., Bell S.P., Gann A., Levine M., Losick R. 2004. Molecular biology of the gene. Benjamin/Cummings, San Francisco, USA.

Chng 9
Di truyn T bo cht
I. S di truyn t bo cht
1. S di truyn ca cc gene lp th Trong s th phn ca thc vt bc cao, mt t bo trng c kch thc ln c nhiu t bo cht phi hp vi nhn ca ht phn khng c t bo cht chung quanh. Do hp t nhn c phn ln t bo cht ca t bo trng. Nu hai b m c thnh phn nguyn liu di truyn trong t bo cht khc nhau th th h con s nhn c nhiu nguyn liu di truyn trong t bo cht ca m. Do s xy ra s di truyn theo th h m. Hin tng di truyn l m c pht hin rt sm Mirabilis jalapa (Correns, 1908), Pelargonium zonale (E.Bauner, 1909). Cc cy c l m c th c nguyn cnh vi l trng khng c chlorophyll. Th nghim: Tp giao gia cy Mirabilis jalapa c nhng cnh khm trng xanh theo cc php lai nh sau: - Th phn cho hoa trn cnh l trng bng ht phn ca hoa trn cnh l xanh lc, cho th h con nhng cy ging c th m c l trng khng c chlorophylle. Cc cy ny cht v khng c kh nng quang hp - Tp giao hoa trn cnh l xanh lc bng ht phn ca hoa trn cnh l trng, tt c th h con c l mu xanh lc bnh thng. - Nu th phn cc hoa ca cnh l m bi phn hoa ca cy xanh lc th i con c cc c th l trng, l m v l xanh lc. - Nu th phn cho ha ca cnh l xanh lc vi phn hoa cy l m th i con gm ton c th l xanh lc. * Gii thch: Trong trng hp ny ngi ta thy nhng cht c s hnh thnh lp th c trong t bo trng s hnh thnh tin lp th, sau hnh thnh lc lp. Ht phn khng c cht c s hnh thnh lp th nn ht phn khng th truyn lc lp c. S khc nhau gia con ci v b m v mt hoc nhiu tnh trng khi tp giao thun nghch chng t c s tham gia ca nguyn liu di truyn trong t bo cht. S di truyn theo h m quy nh s th hin tnh trng ph thuc vo c th m. thc vt Pelargonium zonale c trng hp di truyn theo dng cha. Nu hoa ca cy l m c th phn ca cy l xanh lc th 30% cy lai c l m, 70% l xanh lc. Khi lai hon i cha m th 70% cy

lai l m v 30% l xanh lc. 2. S di truyn ca cc gene ty th 2.1. c im di truyn ca cc gene ty th Theo Mendel, khi tp giao nhng sinh vt lng bi th c s phn ly tnh trng theo ng nh lut ca Mendel v nhng gene trong nhn u nm trn nhim sc th v trong gim phn c phn chia cho cc giao t cng vi nhim sc th. i vi nhng tnh trng trong t bo cht khng c mt h thng phn chia no m nhn nn khng c s phn ly theo mt quy lut nht nh. nm men c mt th t bin c th hnh thnh nhng khun lc petite kch thc nh hn bnh thng, ng knh ch bng 1/2 - 1/2 khun lc bnh thng. Cc t bo to nn khun lc petite c kch thc ging kch thc t bo bnh thng. Nguyn nhn to nn khun lc kch thc nh l do cc t bo t bin petite b hng h thng h hp, tc l nhng enzyme oxy ha trong ty th l cc cytochrom b, c, a, a3 v cytochrom oxydase b ph hy. y l nhng enzyme ca mng trong ty th. Khc vi kiu di, cc t bin petite khng thc hin c phn ng phosphoryl ha sn ra nng lng, v vy tc sinh trng v phn bo ca chng thp hn. ty th ca nm men (Saccharomyces cerevisiae) c 3 kiu t bin ch yu: petite, antR v mit-. Mt v d v ty th l t bin thiu nng h hp nm men. Vo nhng nm 1940, Boris Ephrussi v cs. m t cc t bin c bit nm men.Cc t bin ny c gi l petite, c khun lc nh hn nhiu so vi khun lc hoang di. Theo phng thc di truyn, cc t bin petite chia lm 3 loi khc nhau: - Petite phn ly (Segregation petites): khi lai vi dng hoang di khun lc bnh thng th t l phn ly trong cc nang bo t (ascospore) l 1 khun lc to: 1 petite. - Petite trung tnh (Neutral petites): khi lai vi khun lc to th s phn ly trong nang bo t ch c dng khun lc to bnh thng, th hin s di truyn theo mt cha m (Uniparental) - Petite c ch (Suppressive petites): khi lai to cc nang bo t, mt s mc thnh khun lc to bnh thng, mt s khc to khun lc petite. T l gia khun lc to v nh dao ng nhng c tnh c hiu ca chng, mt s petite c ch ch to th h con khun lc petite. Qua cc petite c ch cho thy c s di truyn ngoi nhn t bo v mt s c s di truyn theo mt cha m.

Khi lai nm men 2 t bo cha m, hai t bo cha m kt hp vi nhau v gp t bo cht nh nhau vo t bo con lng bi. S di truyn ca cc petite trung tnh v c ch c lp vi kiu bt cp th hin r s di truyn ngoi nhn nn c gi l petite t bo cht. Qua nghin cu chng c cc c im kiu hnh nh sau: - Chui chuyn in t ca ty th b sai hng cc petite t bo cht. Do sai hng ny, chng ln men to ATP km nn mc chm. - Khng c sinh tng hp protein cc petite t bo cht. Cc ty th c h thng sinh tng hp ring gm tRNA, cc ribosome khc vi t bo cht. - mtDNA cc t bin petite c bin i ln. Ty th ca tt c cc Eukaryote c mtDNA ring tuy s lng nh, nhng khc vi DNA ca nhn t bo. cc petite trung tnh, mtDNA b mt hon ton, cn cc petite c ch c s thay i ng k t l base so vi mtDNA ca dng khun lc to bnh thng. Nhm cc t bin th hai ca nm men l antR (antR mutants), c kiu hnh khng vi cc khng sinh khc nhau. V d: capR (chloramphenicol resistance) khng chloramphenicol, eryR khng erythromycine, spiR khng spiromycine, parR khng paranomycine v oliR khng oligomycine. Cc t bin ny khi lai (v d eryR eryS) cho t l phn ly khng theo quy lut Mendel, ging nh cc petite c ch nhng s di truyn c khc. Khi cc t bo cha m kt hp, sn phm lng bi l hp t hai cha m cytohet (cytoplasmically heterozygote). Cc diploid ny c th sinh sn v tnh bng mc chi.Trong nguyn phn, qu trnh phn ly t bo cht v ti t hp xy ra v cc t bo con tr thnh eryS hay eryR. Nhm t bin quan trng th ba l mit- (mit- mutants) c pht hin sau cng nh k thut chn lc c bit. Cc t bin ny, tng t cc t bin petite ch c khun lc nh v cc chc nng bt thng ca chui chuyn in t, nhng im khc cn bn l sinh tng hp protein bnh thng v c kh nng hi bin. Nh vy, cc kiu t bin mit- l t bin im. S di truyn cu kiu t bin mit- ging vi kiu antR, c s phn ly t bo cht v s di truyn theo mt cha m trong gim phn. Trong th h con ca nhng t bo thuc khun lc bnh thng, c khong vi phn trm t bo hnh thnh nhng khun lc petite. Nhng t bo khun lc petite lun lun pht trin thnh nhng khun lc petite. iu chng t c s thay i v cu trc di truyn. Ngoi t bin xy ra kiu bo gene ni trn dn n sinh ra nhng khun lc petite, cn c nhng khun lc petite do nhng gene trong nhn quy nh.

Th nghim: Tp giao ca mt ni nm men kch thc khun lc bnh thng vi mt ni c kch thc khun lc petite, th h con hnh thnh khun lc bnh thng. Cn i vi nhng gene trong nhn (gene ade), th s phn ly th h con v nhng gene ny cho t l 1:1, do chng nm trn NST v c chia u cho cc t bo con. y, nguyn liu di truyn trong t bo s c trn ln nhau trong hp t v khi to thnh bo t th mi bo t u nhn c cc gene trong ty th nh nhau, nn chng u c chc nng h hp bnh thng. Th nghim cho thy s di truyn khun lc khng theo quy lut Mendel. 2.2. Hin tng bt dc bo cht c
Ging hu th S to thnh ht phn
Cy bt th c, ht phn mt chc nng T bo cht c di truyn theo dng m
Th h th ba ca lai ngc vi ging bt th c
3/4 gene l ca ging hu Ht to thnh do lai ngc ln th hai
7/8 gene l ca ging hu th c Ht to thnh do lai ngc ln th ba Bt th c trong trng hp ny l kt qu ca di truyn t bo cht
Ht to thnh do lai ngc ln th nht
Ging bt th c
Gene nhn l mt phc hp allele ca b m, t b hu th
Hnh 9.1 S th nghim chng minh s di truyn theo h m ca hin tng bt th c.
Tnh bt dc do nhiu nguyn nhn, bt dc c (khng to phn hoa hay to phn hoa khng c kh nng th tinh) thc vt c cc trng hp sau: - Do gene nhn quy nh, nh gene ms cy ng - Do nh hng ca iu kin mi trng nh m, quang chiu,

kh nng cung cp cht dinh dng khng p ng ng nhu cu sinh l ca cy V d: gene ms cy ng - Do lai xa cng a n cc c th lai khng c ht phn v NST c ngun gc khc nhau khng th tip hp nhau trong gim phn. Nhng hin tng bt dc ny u c ngha hn ch ch c bt dc bo cht c l c vai tr quan trng. l trng hp bt dc ca ht phn bt ngun t t bo cht, cn nhn th c th c iu chnh c nh c th dng cc cy bt dc bo cht c pht huy u th lai cc i tng ng, cao lng, c ci ng... V d: Xt mi quan h gia kiu gene, kiu bo gene v kiu hnh ca bp c s dng trong lai mt tnh m khng cn kh c cy m
STT 1 2 3 4 Kiu gene Rfrf Rfrf RfRf hoc Rfrf RfRf hoc Rfrf kiu bo gene S (bt dc) N (hu dc) N S kiu hnh (ht phn) hng tt tt tt
Vy ht phn ca ng ch b mt hot tnh khi c yu t bt dc trong t bo cht m li thiu thiu gene phc hi hu dc (Rf) trong nhn, alen ca gene ny l rf l gene cng c tnh bt dc. Cy c phc hi hu dc RfrfS cho t th phn th i sau s c 1/4 rfrf c ht phn hng. Nu ly bp ca dng rfrfS th phn ca rfrfN, th phn hoa ca ton b i sau s b hng, cy ny ch cn bp mang nh ci. l cch dng phng php di truyn kh c ng. Khi sn xut ht ging, nhng cy ny mun c ht th phi th phn hu dc ca nhng cy bnh thng. Nu mun dng nhng ht sau ny trng li th cy b phi c kiu gene RfRf v kiu bo gene N hoc S. Trong sn xut ging ng c th dng t hp dng thun dng 1 lm cy m v dng 3 hoc dng 4 ng hp t lm cy b. Nh th s mt cng kh c cy m v ht lai thu c t cy m s c kiu gene Rfrf, kiu bo gene S. Kiu gene ny m bo c s th phn bnh thng lc trng trong sn xut. Bt th c t bo cht ng lin quan n 2 plasmid dng thng S1 v S2. Chng trong ty th cng vi mtADN. Mt trong nhng tnh cht kh hiu ca plasmid ny l chng c th thc hin ti t hp vi mtADN. 3. Hiu qu dng m ln chiu xon v c Trng v phi chu nh hng ca mi trng c th m nhiu hn

c th b. Ngay c khi b tch khi c th m t mt giai on rt sm, chng cng nhn c t bo cht, cc cht dinh dng trong trng t m v nhng nh hng c bit ln hot ng ca gene. Nhng tim nng nht nh ca trng c xc nh trc khi th tinh v trong mt s trng hp chng chu nh hng ca mi trng m bao quanh. S quyt nh trc nh vy do cc gene ca m hn l cc gene ca con, c gi l hiu ng m. S tn ti ca hiu ng m ni chung c chng minh bng phng php lai thun nghch, khi kt qu php lai thun nghch s khc nhau. V d v hiu qu dng m: chiu xon ca v c Limnaea peregra. Chiu xon ny c xc nh bi mt cp alen. D l xon phi, d l xon tri. Cc php lai cho thy D l tri so vi d v chiu xon lun lun c xc nh bi kiu gene ca m.
Th h P
Xon phi Xon tri Xon tri Xon phi
T phi
T phi
Th h F3: 3 xon phi : 1 xon tri
Th h F3: 3 xon phi : 1 xon tri
Hnh 9.2 Hiu qu dng m ln chiu xon v c
Tin hnh php lai thun nghch gia ng hp t DD xon phi v ng hp t dd xon tri: Khi trng bt ngun t c xon phi th F1 l Dd v kiu gene v c c kiu hnh xon phi. Cho cc con c ny t phi th sinh ra th h sau c t l phn li v kiu gene l 1DD : 2 Dd : 1 dd, tt c cc con c thm

ch c con c kiu gene dd u c chiu xon phi. Nhng khi mi con c ca th h ny t phi ring bit th ch c nhng con c kiu gene DD v Dd cho th h sau xon phi, cn nhng con dd mc d c kiu hnh xon phi nhng li cho th h sau xon tri. Ngc li nu dng c dd xon tri lm m th F1 Dd u xon tri ging m. Cho cc c con ny t phi th tt c c th h sau c xon phi v kiu gene ca m chng l Dd. Nu cho mi con ca th h ny t phi th kt qu nhn c nh php lai thun trn, ngha l th h sau c t l phn ly 3 xon phi : 1 xon tri. Nh vy hiu qu dng m ch ko di mt th h v trng hp ny khng th xem l di truyn qua t bo cht bi v y cc c tnh ca t bo cht c xc nh trc bi tc dng ca cc gene trong nhn ch khng phi bi cc gene trong t bo cht. Ni cch khc y c ch di truyn nhim sc th lm bin i t bo cht ca trng trc khi n th tinh.
II. Lp bn ty th v lp th
1. Lp bn gene ca DNA lp th Cc gene ca lc lp Chlamydomonas reinhardii S di truyn lc lp c nghin cu chi tit hn c vi to Chlamydomonas reinhardii. T bo ca vi to ny c mt lc lp ln vi ng knh trung bnh 5 m cha 50 n 80 bn sao ca phn t DNA vng trn di 196 kb. Theo Sager (1975), Chlamydomonas reinhardii c cc t bin trong nhm lin kt gene ca lc lp. Cc t bin c biu hin kiu hnh nh sau: - Mt kh nng quang hp mc c ngoi nh sng v trong ti cn b sung ng kh l acetat - Nhy cm vi nhit cao hoc thp - Tnh khng vi thuc khng sinh hoc c nhu cu c cung cp thuc khng sinh. Tt c cc t bin trn c s di truyn theo mt cha m, c kiu bt cp mt+ (c th coi l dng m). iu ny lin quan n s hnh thnh lc lp trong hp t, bng cch no , ch nhn DNA t lc lp mt+. Nm 1954, R. Sager nghin cu cc t bin khng streptomycin C. reinhardii t dng hoang di nhy cm sm-s. Mt s t bin sm-r c s di truyn nhim sc th vi s phn ly 1 : 1. Tuy nhin mt s t bin c s di truyn khc thng nh sau:

sm-r mt+ sm-s mt- tt c i con u sm-r vi t l 1 mt+ : 1 mt-. sm-s mt+ sm-r mt- tt c i con u sm-s vi t l 1 mt+ : 1 mt-
Hnh 9.3 Bn vng trn ca cpDNA Chlamydomonas
Hnh 9.4 Bn DNA chloroplast ca Marchantia polymorpha IRA v IRB l nhng trnh t o ngc (theo K.Umesono v H.Ozeki, 1987)
B
Nh vy y khi c s hon i cha m trong lai, th h con u c kiu hnh streptomycin ca mt+. S truyn th tnh trng ny c gi l s di truyn theo mt cha m. Sager coi mt+ nh dng m v trng hp trn ging nh di truyn theo dng m. Cc kiu bt cp mt c t l phn ly ca gene trong nhn l 1 : 1. Trong t hp lai mt+ sm-r mt- sm-s c khong 0,1% th h hp t con mang c sm-r v sm-s. Cc hp t nh vy gi l hp t hai cha m cytohet (cytoplasmically heterozygote).

Tn s cc cytohet c th tng ln 40-100% th h hp t con nu mt+ c chiu tia t ngoi trc khi lai. Chlamydomonas, cc hp t 2 cha m c dng lm im xut pht cho tt c cc nghin cu v s phn ly v ti t hp ca cc gene lc lp. Trn c s nhiu t hp lai, R. Sager nu ra bn vng trn ca cpDNA vi cc gene tng ng. 2. Lp bn gene ca DNA ty th * Lp bn b gene ty th ca nm men C c c phng php khc nhau xy dng bn b gene ty th: - Lp bn ti t hp (recombination mapping): nm men s phn li t bo cht v ti t hp xy ra trong qu trnh mc chi cytohet lng bi. S phn li v ti t hp c th pht hin trc tip cc dng lng bi mc chi hay quan st sn phm gim phn khi chi c kch thch to bo t. V d khi lai eryRspiR erySspS c th nhn c cc kiu b bn vi s lng nh sau: eryRspiR 63 b bn S S ery spi 48 b bn S R ery spi 7 b bn R S 1 b bn ery spi S R Cc kiu gene ery spi v eryRspiS l cc dng ti t hp. - Lp bn bng phn tch petite: Mt s k thut lp bn c hiu qu da trn s phi hp c 3 loi t bin petite, antR v mit-. Phn ln cc tip cn ny da vo s mt on ca mtDNA ca cc t bin petite. S kt hp cc kiu phn tch di truyn c bit vi k thut ti t hp DNA dn n xy dng bn di truyn hon chnh ca mtDNA. - Lp bn bng phn tch enzyme ct gii hn: cc enzyme ct gii hn (xem chng 10) l cng c hu hiu mi phn tch di truyn. N c s dng khng nhng phn tch mtDNA ca nm men, m c mtDNA ca bt k sinh vt no min chit tch v tinh sch c DNA. * Bn mtDNA ca nm men v ngi Vic xy dng bn mtDNA hon chnh ca nm men v ngi l nhng thnh tu ng k ca nghin cu di truyn t bo cht. - Mt s trnh t c codon khi s v codon kt thc cui nhng cha bit chc nng

- mtDNA ca ngi rt t d tha
Phin m ca si nh
Phin m ca si nng Nhng trnh t n m ha cho amino acid trn tRNA mtDNA ca ngi
Kt qu t bin tRNALys vng MERRF
Hnh 9.5 Bn mtDNA ca ngi (Theo Larson v Clayton, 1995)
III. Di truyn hc phn t cc bo quan

1. Cc b gene lp th (cpDNA) L bo quan c kh nng t ti sinh t bo thc vt. S phn chia ca cc bo quan ny v v cc t bo con trong phn bo l khng u nh s phn chia ca nhim sc th trong nguyn phn v gim phn. Chng c s lng ln v phn chia ngu nhin v cc t bo con nn mi t bo c th cha nhiu hoc t lc lp. DNA ca lc lp c k hiu l cpDNA (Chloroplast DNA). B gene ny dng DNA vng trn, thng di hn DNA ca ty th 8-9 ln. Trong lc lp cn tm thy b my sinh tng hp protein khc rt nhiu vi h thng trong t bo cht ca eukaryote nhng ging vi b my sinh tng hp protein ca prokaryote. Mc d s di truyn ca lc lp c pht hin rt sm, nhng trong mt thi gian di s hiu bit chi tit v cc gene ca lc lp khng c bc tin ng k. Cc nghin cu phn t gp phn ch yu cho s phn tch chi tit cc gene cc bo quan. Ngoi cc nghin cu Mirabilis jalapa v Chlamydomonas, bn chi tit cpDNA ca thc vt Marchantia polymorpha c xy dng.

cpADN in hnh di khong 120-200 kb ty loi thc vt. Marchantia, kch thc phn t l 121 kb. Trn cpDNA ca Marchantia c tt c 136 gene gm 4 loi m ha tng hp rRNA, 31 loi m ha tng hp tRNA v khong 90 gene tng hp protein. Trong s 90 gene m ha tng hp protein, c 20 gene m ha tng hp enzyme cho quang hp v chui chuyn in t. Cc gene m ha cho cc chc nng dch m chim khong mt na b gene ca lc lp v bao gm cc protein v cc RNA cn thit cho dch m bn trong lc lp. Thc t DNA ca lc lp, ty th v nhn t bo c s phi hp cht ch trong vic to ra cc tiu phn ca nhng protein c s dng bn trong lc lp. Ribulose-1,5-biphosphate carboxylase/ oxygenase l enzyme di do nht ca lc lp. N xc tc 2 phn ng cnh tranh nhau, c nh CO2 v bc u tin ca quang h hp (photorespiration) vi s to ra glycolate. Enzyme gm 8 tiu phn ln LS (large unit) ging nhau v 8 tiu phn nh ging nhau c m ha tng ng bi cc gene ca lc lp v nhn t bo. Tiu phn ln LS mang trung tm xc tc, cn vai tr ca cc tiu phn nh cha r. Gene LS nm trn cpDNA ca mt s thc vt nh bp, Chlamydomonas reinhardii, thuc l, Euglena... Trong tt c cc trng hp, gene LS hin din 1 bn sao cho 1 DNA ca lc lp. Ngc li, cc gene ca tiu phn nh c tm thy cc trnh t DNA ca nhn t bo vi s bn sao t. 2. Cc b gene ty th (mtDNA) Bo quan ty th c tt c cc t bo ca eukaryote. B gene ca ty th c k hiu l mtDNA (Mitochodrial DNA). mtDNA m ha cho s tng hp nhiu thnh phn ca ty th nh h thng 2 loi rRNA, 22-25 loi tRNA v nhiu loi protein c trong thnh phn mng bn trong ty th. Trong khi , phn ln protein ca ribosom ca ty th th do cc gene trong nhn xc nh. B gene ca ty th c hai chc nng ch yu: - M ha cho mt s protein tham gia chui chuyn in t - M ha cho h thng sinh tng hp protein gm mt s protein, tt c cc tRNA v c 2 loi rRNA. Tuy nhin trong c hai trng hp, nhng cu phn cn li ca h thng c m ha do cc gene nhn v c dch m bo tng (cytosol) ri chuyn vo ty th. Vit Nam c cng trnh phn tch mtDNA 50 ngi Vit dn tc Kinh, pht hin c cc hnh thi khc nhau khi ct mtDNA bng 6

enzyme hn ch. Nh vy, vic nghin cu cc gene ca ty th cho thy t bo eukaryote khng lc lp c t nht 2 h thng sinh tng hp protein c lp tng i nhng lun hp tc cht ch vi nhau. cc eukaryote c lc lp th 3 h thng sinh tng hp protein c lp tng i nhng hp tc vi nhau. C 2 bo quan ty th v lc lp tham gia trc tip vo chuyn ha nng lng ca t bo. Di truyn t bo cht l hin tng di truyn do cc gene nm trn nhim sc th ngoi nhn quy nh.
Cu hi v Bi tp
1. Hy nu cc kiu t bin ca ty th. 2. Trnh by chng minh thc nghim v di truyn lp th. 3. So snh s ging nhau v khc nhau ca mtDNA v cpDNA. 4. Nu c ht bp t dng bt thu c, lm sao xc nh s bt th do gene trong nhn hay ngoi nhn? 5. Nu cc c im ring ca di truyn ngoi nhim sc th v nu cc sai khc c bn so vi di truyn ca gene nhn. 6. Hy phn bit hiu qu dng m vi s di truyn ngoi nhn. 7. Hy phn bit cc loi t bin petite nm men. 8. Cho 2 dng bp bt dc c: mt dng bt dc bo cht, mt dng bt dc do gene nhn di truyn theo Mendel. Hy trnh by phng php xc nh hai dng bt dc trn. 9. Mt con c loi Limnaea peregra c v xon tri qua t phi cho tt c th h sau xon phi. Hy xc nh genotype ca con c ny. Nu th h sau cho t phi ring r, hi chiu xon v ca cc con c th h ny nh th no? 10. T l tin ha DNA ca ty th c tnh bng s bin i 1 nucleotide ca 1 ty th trong thi gian 1.500 n 3.000 nm. DNA ty th ngi c khong 16.500 nucleotide. Hi t l bin i ca 1 nucleotide trong 106 nm l bao nhiu?
Ti liu Tham kho

Ting Vit Phm Thnh H. 2000. Di truyn hc. NXB Gio Dc.

L nh Lng, Phan C Nhn. 1998. C s di truyn hc. NXB Gio dc. Phan C Nhn, Nguyn Minh Cng, ng Hu Lanh. 1999. Di truyn hc. NXB Gio dc Hong Trng Phn. 1995. Di truyn hc phn t. Trung tm o to T xa, i hc Hu Ting Anh Anthony J. F. Griffiths, Susan R. Wessler, Richard C. Lewontin, William M. Gelbart, David T. Suzuki, Jeffrey H. Miller. 2004. An introduction to genetics analysis. W.H. Freeman Publishers. Harlt D.L., Jones E.W. 1998. Genetics - Principle and analysis. Jone and Bartlett Publshers, Toronto, Canada. Stansfield W.D. 1991. Schaums outline of theory and problems of genetics. McGraw-Hill, Companies, Inc., United States of America. Vu Trieu An. 1999. Mitochondrial DNA Polymorphism in the Vietnamese population. Eur. J. of Immunogenetics, 26: 471- 422. Watson D.J, Baker T.A., Bell S.P., Gann A., Levine M., Losick R. 2004. Molecular biology of the gene. Benjamin Cummings, San Francisco, United States of America.

Chng 10
i cng v Cng ngh DNA Ti t hp

Vic tinh ch enzyme ct gii hn u tin (1970) v s dng n to ra cc phn t DNA ti t hp u tin trong ng nghim (1972-1973) l nn tng cho s ra i ca k thut di truyn (genetic engineering) v cng ngh DNA ti t hp (recombinant DNA technology). Chnh s pht trin nhanh chng ca lnh vc ny khng nhng a li khi lng tri thc khng l v cu trc v c ch hot ng ca cc gene, cc b gene prokaryote v eukaryote m cn tr thnh lc lng sn xut trc tip ca x hi: to ra hng lot cc ch phm y-sinh hc hu ch t cc t bo vi khun, nm men; to cc ging sinh vt mi... gp phn gii quyt nhng vn thc tin t ra trong y hc v trong cng tc chn to ging. Trong chng ny chng ta s tm hiu cc c tnh ca enzyme ct gii hn, cc nguyn l c bn ca k thut DNA ti t hp v mt s ng dng ca lnh vc cng ngh ny.
I. Cc cng c chnh ca k thut to dng DNA ti t hp

1. Cc enzyme ct gii hn 1.1. Enzyme ct gii hn l g? Enzyme ct gii hn (restriction endonuclease) hay gi tt l enzyme gii hn (restrictase) l loi enzyme c kh nng nhn bit on trnh t nucleotide c hiu trn cc phn t DNA v ct c hai si DNA b sung ti cc v tr c th. 1.2. Vai tr ca cc enzyme ct gii hn T 1953 ngi ta pht hin thy rng, khi a DNA ca mt ni vi khun E. coli ny vo t bo thuc mt ni khc thng th DNA c a vo, gi l DNA ngoi lai hay DNA l, mt hn hot tnh di truyn v hu nh bao gi cng b phn ct thnh cc on ngn. Ch trong mt s t trng hp DNA l mi khng b phn ct v do n c th ti bn trong t bo ch. iu chng t DNA l c sa i bng cch no di s kim sot ca t bo ch. Cc hin tng ni trn xy ra ch yu khi cc th thc khun (phage) xm nhim cc t bo vi khun. Cho n u thp nin 1970 ngi ta mi bit r rng cc t bo vi khun l nhng h thng cha c hai loi enzyme: cc enzyme sa i v cc enzyme ct gii hn. Chng u c i tng nhn bit l cc on trnh t ca DNA vt ch v DNA ngoi lai, nhng c vai tr khc nhau. C th, cc enzyme sa i (methylase) ng vai tr bo v DNA vt ch

bng cch gn thm nhm methyl (-CH3) mt s base nht nh trong on nhn bit (recognition sequence) hay on ch (target sequence). Hin tng methyl ho (methylation) ny thng xy ra i vi adenine v bin i n thnh N-6 methyladenine. Trong khi , cc enzyme gii hn li ng vai tr v hiu ho hot tnh di truyn ca cc DNA l bng cch phn ct cc v tr c th chng no n cha c sa i cho ging vi DNA vt ch. Nh vy, cc enzyme gii hn ng vai tr l hng ro bo v t nhin ca cc vi khun nhm chng li s xm nhp ca cc phage l. 1.3. Tnh cht chung ca cc enzyme gii hn Trc tin, cn lu rng cc enzyme gii hn ch pht hin thy cc vi khun m khng c cc eukaryote. V vy, tn gi ca cc enzyme gii hn thng dng l tn h thng, c biu th bng ba hoc bn ch ci vit tt ca vi khun m t enzyme c chit xut. Ch ci u tin c vit hoa ch chi (genus) v hai ch ci tip theo vit thng ch loi (species), v khi cn thit thm ch ci th t ch ni hoc chng (strain, type). Ngoi ra, phn bit cc enzyme cng mt ni ngi ta dng s La M km theo sau tn h thng (xem bng 10.1).
cc on DNA ni cc u dnh
u dnh
u dnh
+ DNA ligase
DNA ti t hp
Hnh 10.1 Enzyme gii hn (EcoRI) ct cc DNA khc nhau, nh c th kin to phn t DNA ti t hp in vitro (bng enzyme DNA ligase).
Tnh cht quan trng nht ca cc enzyme gii hn l tnh c hiu v tr, ngha l chng c th nhn bit on trnh t DNA c th ct v tr xc nh. Tu theo v tr ct so vi on nhn bit m chia ra hai loi:

loi I bao gm cc enzyme gii hn ct bn ngoi phm vi on nhn bit v loi II bao gm cc enzyme ct c hiu bn trong on nhn bit. y chng ta ch xt cc enzyme gii hn loi II vn c xem l cng c hiu nng (ging nh con dao m tinh vi) cho php thao tc trn cc gene trong k thut DNA ti t hp (hnh 10.1). c trng ni bt ca cc on ch l c kch thc ngn, 4-8 cp base, v c tnh i xng xui ngc (palindrome). Nhn chung, cc enzyme gii hn khc nhau c hai kiu ct sau y: ct lch v ct thng. Vi kiu ct lch tc l cc v tr ct trn hai si ca DNA si kp l so le, to ra cc on DNA c cc u si n gm mt s base b sung gi l cc u dnh (cohesive/sticky ends). Cc enzyme gii hn nh th c vai tr to ln trong vic kin to DNA ti t hp in vitro (hnh 10.1). in hnh y l EcoRI v BamHI (bng 10.1). Vi kiu ct thng, tc ct cng v tr trn c hai si ca DNA si kp, do to ra cc on DNA c cc u bng (blunt ends); v d, SmaI...(bng 10.1). Cc enzyme gii hn khc nhau c on ch ging nhau, mc d v tr v kiu ct c th ging hoc khc nhau, gi l cc enzyme gii hn tng ng (isoschizomers); v d, SmaI v XmaI (bng 10.1).
Bng 10.1 Cc trnh t nhn bit v v tr ct ca cc enzyme gii hn c chn lc (mi tn ch v tr ct; cc trnh t y c ch ra trn si 5'3') Ngun vi sinh vt Arthrobacter luteus Bacillus amyloliquefaciens H Escherichia coli RY13 Haemophilus influenzae Rd Haemophilus influenzae Rd Nocardia otitidis-caviarum Providencia stuartii Serratia marcescens Sb Xanthomonas malvaccarum Tn enzyme AluI BamHI EcoRI HindII HindIII NotI PstI SmaI XmaI Trnh t nhn bit AGCT GGATCC GAATTC GTPyPuAC AAGCTT GCGGCCGC CTGCAG CCCGGG CCCGGG
2. Cc vector thng dng trong k thut di truyn 2.1. DNA ti t hp l g? DNA ti t hp (recombinant DNA) l phn t DNA c to ra trong ng nghim bng cch kt hp cc DNA t cc ngun (loi) khc nhau, theo mt quy trnh k thut nht nh, gi l k thut ti t hp DNA. Thng thng mt phn t DNA ti t hp bao gm mt phn t DNA c bn cht l plasmid hoc phage nguyn vn gi l vector (th ti) v mt on DNA t ngun khc mang mt gene hoc yu t iu ha mong

mun c cho xen vo; n c gi l DNA ngoi lai (foreign DNA). 2.2. Hai loi vector thng dng Vector l phn t DNA c kch thc b hoc va phi, ng vai tr l vt trung gian mang truyn on DNA ngoi lai nghin cu vo trong t bo th nhn (t bo kh bin) bng con ng bin np (transformation) hoc ti np (transduction). C hai loa vector thng dng l cc plasmid hoc cc phage. Cc plasmid vi khun (hnh 10.2) c s dng rng ri hn c, bi v chng c cc c im sau: (i) c kh nng xm nhp vo t bo vt ch m vn hot ng (ti bn, biu hin gene) bnh thng; (ii) c trng lng phn t thp nn d dng tinh chit; (iii) s bn sao trong mi t bo vi khun thng kh cao; v (iv) c bit l, mt s plasmid c cha cc gene khng thuc tin li cho vic theo di v pht hin s c mt ca plasmid ti t hp trong vi khun ch.
cc khi im ti bn
Hnh 10.2 Cc plasmid c cha khi im ti bn (ori), cc im ct ca mt s retrictase v cc gene khng ampicillin (AmpR), kanamycin (KanR).
Trong s cc phage dng lm vector th phage lambda () c nhiu u th nht, bi l phn gia ca b gene c cha mt s gene khng quan trng v khng lin quan vi s ti bn ca n, nn thun li cho vic xen on DNA mong mun vo y. Cc phage khng cha cc gene khng thuc cho nn vic theo di phage ti t hp c xc nh da vo cc vt tan dng tnh (positive plaques) trn nn vi khun. 3. Thit lp phn t DNA ti t hp in vitro

3.1. Phng php s dng cc u dnh Bt k on DNA no nu c ct bi cng mt loi enzyme gii hn (v d, EcoRI) cho cc u dnh th c th dnh lp li vi nhau v c ni bi DNA ligase (hnh 10.3). Phng php thnh lp phn t DNA ti t hp kiu ny ln u tin c a ra bi J.Mert v R.Davis nm 1972 bng thc nghim trn cc virus. V sau , ln u tin nm 1973, H.Boyer v nhm nghin cu ca S.Cohen to ra c phn t DNA ti t hp gm vector l plasmid nh pSC101 ca E. coli v DNA ''ngoi lai'' l mt plasmid khc. Chnh s kin ny t nn mng v m ra trin vng to ln cho k thut DNA ti t hp sau ny.
DNA s c xen vo
DNA c ct vi EcoRI
Cc u dnh
Ti t hp DNA + DNA ligase
Ni sai
DNA ti t hp
Ni sai
Hnh 10.3 Hai phn t DNA khc nhau c ct bi cng mt enzyme gii hn th c th ni vi nhau nh xc tc cu DNA ligase.
3.2. Phng php ni trc tip hoc tng hp cc u b sung i vi cc on DNA c to ra bng cch x l enzyme gii hn ct thng nh HindII chng hn, th vic ni cc on DNA c u bng c to ra c th thc hin theo hai cch sau: Ni trc tip bng DNA ligase ca phage T4 hoc tng hp thm cc u dnh vo cc u 3' mt s nucleotide b sung bng cch s dng cc enzyme end transferase, ri sau cc on DNA nh th s c ni vi nhau bi DNA ligase ca vi khun. C s ca phng php kt hp DNA ny c thc hin ln u tin gia DNA ca virus SV40 vi DNA ca phage bi L.Lobban v

D.Kaiser (1972), v D.Jackson v P.Berg (1972)
II. To dng gene hay DNA ti t hp

1. Nguyn tc chung V nguyn tc, k thut DNA ti t hp hay to dng (cloning) gm cc bc chung nht nh sau: (1) Tch chit v tinh sch DNA thuc cc ngun khc nhau (gm vector v DNA mang gene mong mun); (2) to ra phn t DNA ti t hp in vitro; (3) a phn t DNA ti t hp vo trong t bo nhn, thng l E. coli hoc nm men. Hnh 10.4 m t mt quy trnh k thut n gin nh th. Tuy nhin, trn thc t, s phc tp l bc (4), pht hin v phn lp cc dng DNA ti t hp c hiu.
DNA ngoi lai Cc on ct bi enzyme gii hn Enzyme gii hn Ni cc u dnh
Plasmid ti t hp
Bin np
T bo ch E. coli
T bo c bin np
Hnh 10.4 Mt quy trnh bin np DNA ti t hp vi vector l plasmid, enzyme gii hn EcoRI, enzyme ni - DNA ligase, v t bo nhn l E. coli.
Trong t bo ch, phn t DNA ti t hp c th biu hin gene mong mun (cho sn phm protein) hoc ti bn c lp nhiu ln to ra hng lot bn sao ca n, v khi t bo ch phn chia s ko theo s to dng phn t (molecular cloning). Mt khc, do tc phn chia rt nhanh ca cc vi khun nn c th to hng triu bn sao mong mun trong mt thi gian ngn. V th nh khoa hc c th tch dng bt k mt gene no

dng cho nghin cu hoc cho sn xut trn quy m cng nghip mt s lng ln cc protein vn l nhng ch phm y-sinh hc no . 2. Quy trnh to dng gene ti t hp By gi ta xt mt quy trnh k thut to dng m vic pht hin DNA ti t hp da trn kh nng khng thuc do vector plasmid mang li. Bc 1: Tinh chit DNA Gi s tinh chit c plasmid c cha hai gene khng ampicillin v tetracyclin, k hiu l AmpR v TetR; v cng gi thit rng gene TetR c cha im ct ca EcoRI, v phn t DNA ngi c mang gene insulin. Bc 2: Kin to phn t DNA ti t hp in vitro Trc tin, dng enzyme gii hn u dnh EcoRI (xem bng 10.1) ct vng plasmid ti gia gene TetR v cho ct DNA ngi, trong s cc on b ct c mt on mang gene insulin. Sau em trn ln hai loi DNA trn trong ng nghim vi DNA ligase. Kt qu l c th xy ra ba trng hp: (1) Plasmid t ni li thnh mch vng nh lc u; (2) on DNA t ni li thnh mch vng; v (3) Plasmid ti t hp c mang gene insulin, v c th mang mt on DNA khng phi gene . Bc 3: Bin np v pht hin dng DNA ti t hp chung a cc DNA c x l vo cc t bo E. coli. Nu phn t c kch thc ln ngi ta phi x l vi khun 'th nhn' bng chlorid calcium (CaCl2) lm cho mng tr nn thm c d dng. Sau em cy ring r cc vi khun trn mi trng c ampicillin v theo di. + Nu c xut hin khun lc (cc vi khun trong cng mt khun lc th thuc mt dng v chng bt ngun t mt vi khun ban u), chng t vi khun c mang gene AmpR, tc l chng c mang plasmid ban u (trng hp 1) hoc plasmid ti t hp (trng hp 3). Ngc li, nu ch cy khng xut hin khun lc, chng t vi khun mang DNA t ni (trng hp 2). + K , em cy ring r cc vi khun thu c sang mi trng c tetracyclin. Nu c xut hin khun lc, chng t vi khun c mang gene TetR nguyn vn (trng hp 1). Nu khng c khun lc, chng t vi khun em cy c mang DNA ti t hp (trng hp 3); v gene TetR b bt hot do on DNA xen vo. Bng cch theo di nh vy cho php xc nh c dng vi khun mang DNA ti t hp, nhng vn cha bit c u l cc dng c hiu, ngha l c mang gene insulin. Hnh 10.5 di y m t mt quy trnh n gin v to dng vi khun mang gene insulin ngi.

gene TetR gene AmpR DNA ngi gene insulin
cc u dnh
Lai ho + DNA ligase Bin np T bo vi khun To dng Nui cy Dng

Ch c cc vi khun cha DNA ti t hp sinh trng c
NST vi khun
t cc vi khun ln mi trng c b sung ampicillin
Tinh chit DNA
Hnh 10.5 S th nghim to dng vi khun mang DNA ti t hp, y l gene insulin ngi.
Bc 4: Chn dng DNA ti t hp c hiu V nguyn tc, trong c triu php th mi c mt t bo mang gene mong mun. Vi trng hp trn y chng hn, ngi ta c th s dng phng php min dch hc bng cch dng khng th chng li protein c sinh ra bi dng vi khun tng ng (tc huyt thanh tm gene khng insulin). Ni chung, tm dng lai c hiu ngi ta s dng cc mu d l mRNA hoc rRNA c hiu. Chng hn, trong trng hp nu cn chn dng lai mang on mRNA c th, ngi ta em cy u cc dng vi khun c cha DNA ti t hp ln trn mt thch ca hp petri cha mi trng nui cy. Sau ng du ln mng lc nitrocellulose, v thu c bn sao. Vic x l bn sao bng NaOH s lm cho cc t bo vi khun tan v ti ch (in situ), v cc DNA thot ra t chng s b bin tnh (cc si n tch ri nhau) v dnh vo mng lc. Sau em nhng

mng lc ny vo mu mRNA tng ng c tinh khit v nh du phng x (P32); mu RNA ny c gi l vt d phng x (radioactive probe). Nu dng no c cha DNA m ho cho mRNA th s xy ra hin tng lai gia mRNA v vng si n tng ng trn DNA . Sau khi loi b cc mRNA khng lai c, ngi ta t mt ming phim nh ln trn mng lc; nhng vt nh xut hin trn nh phng x t ghi cho thy v tr ca dng mang DNA b tr vi mu RNA. T y c th tch ring cc dng lai c hiu s dng cho cc nghin cu tip theo.
Sinh trng qua m
T bo + pUC19 T bo + pUC19 T bo khng + on xen c bin np Mi trng + ampicillin + X-gal Cc khun lc trng: pUC19 + on xen Cc khun lc xanh: ch c pUC19
Cc dng khng ampicillin Vt d phng x
(a)
(b)
Mng
Cc dng chn lc Mng Phim tia X
(c)
Hnh 10.6 Xc nh cc dng vi khun mang plasmid c xen mt on DNA (gene) c hiu bng vt d phng x l mRNA - sn phm ca n.
Hnh 10.6 minh ha mt cng on ca quy trnh th nghim DNA ti t hp vi khun E. coli khi s dng mi trng nui cy c b sung ampicillin i vi ba kiu t bo: t bo c mang plasmid ti t hp, t bo ch mang plasmid pUC19 khng ti t hp, v t bo khng bin np c (hnh 10.6a). Qua m sinh trng, cc t bo no c mang plasmid ti t hp v plasmid khng ti t hp s mc thnh cc khun lc (mu sc tng ng y l trng v xanh; hnh 10.6b). Sau khi chn ra cc dng c xen plasmid ti t hp, a ln mng lc v cho tin hnh lai ha gia RNA v DNA (gene) ca n bng cc vt d phng x nh ni trn. Sau a sn phm lai phn t ny ln tm phim X quang nh v

gene quan tm t dng ti t hp (hnh 10.6c). 3. Tng hp v to dng cDNA i vi trng hp cn cho biu hin mt gene l (sn xut mt protein) mong mun trong vi khun, ngi ta c th tng hp gene ca n da trn khun mu mRNA v enzyme phin m ngc tinh ch t cc virus RNA (xem chng 5). Sau cho xen gene ny vo plasmid, ri em cy vo vi khun v xc nh cc dng cDNA c hiu. y ta ch xt hai bc u: Bc 1: Tng hp cDNA Nh bit, cc mRNA eukaryote u c ci ui poly(A) u 3'. Chnh trnh t ny to iu kin thun li cho vic tng hp si DNA b sung, cDNA. Khi em trn ln cc on ngn gm cc nucleotide thymine (oligodT) vi mRNA ny s xy ra s lai ho gia n vi vng ui mRNA. on oligo(dT) lm mi cho enzyme phin m ngc (reverse transcriptase) tng hp si cDNA m sn phm l si kp lai RNAcDNA. u 3' ca si cDNA c tng hp c ci 'chp' (tng t u 5' ca mRNA). Tip theo, bng cch x l vi NaOH, si mRNA b loi ra; k ci 'chp' u 3' ca cDNA li lm mi cho DNA polymerase I tng hp si th hai dc theo si khun vn c ca n. Sn phm cDNA by gi cn mang ci 'vng' si n. Sau , ci vng ny c ct b bng cch x l vi nuclease S1 to ra cDNA si kp. Bc 2: Xen on cDNA vo plasmid xen on cDNA vo plasmid ngi ta c th dng enzyme end transferase gn thm ''ui'' homopolymer (v d, CCCC....) vo cc u 3' ca cDNA. V plasmid sau khi c m vng, cng phi lp thm u 3' nhng trnh t tng ng l GGGG... cng vi enzyme trn. Tuy nhin, cch ph bin hn c l gn thm vo c hai 'u bng' ca si kp cDNA ny bng cc oligonucleotide gm 8-10 cp base, nh xc tc ca DNA ligase T4. Sau , dng enzyme gii hn thch hp ct ''on ni'' ny to ra cc u dnh. ng thi ct plasmid bi cng mt enzyme ti gene TetR. Hai DNA ni trn c ni vi nhau bng DNA ligase to ra plasmid lai.
III. Cc phng php biu hin cc gene c to dng

Mt s cc vector c s dng trong cc th nghim to dng ti t hp (nh cp) vn c th c s dng nh l nhng vector biu hin (expression vectors). Cc vector ny c th sn sinh ra cc sn phm protein ca cc gene c to dng. Chng hn, cc vector pUC v pBS c xen vo DNA di s kim sot ca lac promoter, vn nm pha

trc so vi v tr to dng phc (multiple cloning site). Nu nh mt on DNA c cho xen c mt trong cng khung c m th n lm gin on gene lacZ', s sinh ra mt protein dung hp (fusion protein). N s c mt phn trong trnh t ca protein beta-galacyosidase ti u amin v trnh t protein khc na vn c m ha trong DNA c xen vo, u carboxyl ca n. Tuy nhin, nu ta quan tm ti s biu hin cao ca vector c to dng, th cc vector chuyn biu hin thng hot ng tt hn. C hai yu t in hnh cn thit cho s biu hin gene c hot tnh: mt promoter mnh v mt v tr bm ca ribosome m bao gm lun c trnh t Shine-Dalgarno nm gn codon khi u AUG. Trn thc t, ngi ta s dng cc vector biu hin c cc promoter mnh (expression vector with strong promoters), chng hn nh promoter ca operon tryptophan. N to thnh c s cho nhiu vector biu hin k c ptrpL1. Ngoi ra, ngi ta cn s dng cc vector biu hin dng cm ng (inducible expression vectors). Trng hp ny thng tin li ch, n gi cho mt gene c to dng trng thi ng cho ti khi ta sn sng cho n biu hin. Mt l do l ch, cc protein ca eukaryote c sn sinh mt s lng ln vi khun c th gy c. Ngay c cc protein vn khng c thc s, chng cng c th c to ra nhiu n mc gy ri lon s sinh trng ca vi khun... Promoter ca operon lactose (lac promoter) l vector biu hin kiu cm ng n mt mc no , c l l vn gi bt hot cho ti khi c kch hot bi cht cm ng allolactose hoc bng cht tng hp tng t ca n l IPTG. Tuy nhin, s biu hin vn km bi cht c ch lac l khng y hon ton, v s biu hin no ca gene c to dng vn c th pht hin c ngay c khi khng c mt cht cm ng. Mt cch xoay quanh vn ny l cho biu hin gene mong mun trong mt plasmid hay phagemid m n mang c gene lacI ca ring n, nh l plasmid pBS chng hn. By gi chng ta th tm hiu mt phng php sn xut insulin ngi bng con ng tng hp DNA v cho biu hin gen E. coli. Trc tin cn lu rng, thc hin c iu ny ngi ta phi da trn thnh tu mi nht t vic nghin cu cu trc chi tit v qu trnh tng hp cc ch tit insulin t tuyn tu vo mu. Ni vn tt th sn phm s cp ca qu trnh dch m t phn t mRNA hon chnh l preproinsulin gm on peptide "tn hiu dn u" v cht tin thn ca insulin l proinsulin; on pre- b tch b trong qu trnh tng hp. Proinsulin c ch tit l phn t gm ba on A, B v C lin nhau trong mt cu trc "hnh quai" c ba cu disulfur; khi on peptid C (33 amino acid) b ct b bi enzyme

c th trong cc ti ca t bo tuyn ty s to ra cc sn phm insulin c hot tnh. Phn t insulin gm hai chui polypeptid A (21aa) v B (30aa) ring bit c duy tr cng nhau bi hai cu disulfur. T y ta c th hnh dung qu trnh tng hp gene insulin nhn to (cDNA) v cho sn xut hormone ny E. coli nh sau. Trc tin, dng mRNA ca proinsulin lm khun tng hp y mt DNA si kp bng con ng phin m ngc nh trnh by trc. Sau lp thm b ba khi u ATG nhn to (m ho amino acid u chui polipeptide) vo u 5' bng phng php ho hc. Tip n, cho n kt hp vi mt phn ca operon lactose (gm mt on ca gene galactosidase v ton b promoter) ca E. coli n c th hot ng c trong t bo th nhn. Sau gene "lai" ny c xen vo plasmid pBR322 ( y khng i su vo cc chi tit k thut, chng hn nh s dng cc on ni, linker, v cc enzyme gii hn). Khi a plasmid lai ny vo vi khun E. coli, n s sn sinh ra cc protein lai gm mt on peptide ca -galactosidase ni lin vi phn t proinsulin qua gc methionine (MET). Sau khi phn lp protein ny v x l in vitro bng cyanogen bromide th gc MET b ct b (ko theo phn enzyme ca vi khun l -galactosidase tch ra) v thu c proinsulin nguyn vn. Cui cng, nh x l vi enzyme thch hp, on peptid C gia proinsulin c tch ra v c th thu c insulin dng tinh khit. Cng cn lu rng, hin nay ngi ta to ra c cc dng vi khun v cc chng nm men mi c hiu c kh nng sn xut insulin trn quy m cng nghip vi ch s insulin cao, v c bit l, cc chng ny c th trc tip bi xut sn phm c hiu vo mi trng nui cy. Trong trng hp , cc t bo chuyn sn xut insulin vn c duy tr v ti s dng vi hiu qu kinh t cao.
IV. ng dng ca cng ngh DNA ti t hp

1. Cng ngh DNA ti t hp vi vic nghin cu b gene Vic tch dng ti t hp cho php nhn c mt s lng ln bt k gene hoc vng iu ho no tin hnh phn tch trnh t nucleotide, xc nh cc vng chc nng v ch ra cc c ch hot ng ca chng. Nh a li nhng hiu bit mi v t chc v hot ng ca cc b gene prokaryote (nh cc khi im ti bn, cc vng iu ho phin m, cc gene nhy v.v.) v cc b gene eukaryote (nh cc centromere, telomere, cc gene phn on, cc gene gi, DNA lp li v.v.) nh c tho lun cc chng trc.

DNA ngi vector YAC
DNA cn to dng v DNA plasmid c ct bi cng enzyme gii hn
ct tng phn u dnh cc on NST ln u dnh
Cc plasmid cha cc on xen khc nhau
u dnh Ti t hp + DNA ligase Nhim sc th nm men nhn to xen DNA ngi
vi khun c bin np vi hn hp hoc cc plasmid nui cy vi khun ch mi dng l mt 'volume' trong th vin gene
lln ti 1.000.000 bp Bin np vo t bo nm men
Dng
(a) (b) Hnh 10.7 Xy dng th vin gene hay th vin b gene (gene or genomic library) bng cc th nghim to dng plasmid ti t hp vi khun (a) v s dng nhim sc th nm men nhn to, YAC (b).
Bng cc th nghim to dng plasmid ti t hp kinh in vi khun E. coli, v bng cch ci tin s dng nhim sc th nm men nhn to ny (yeast artificial chromosome = YAC; hnh 10.7), ngi ta thnh lp cc th vin gene (gene library) hay th vin b gene (genomic library) trn c s tin hnh lp bn vt l (physical map) v xc nh trnh t DNA b gene ca nhiu sinh vt khc nhau, k c b gene ngi (NHGRI 2005). Nu nh vo nm 1977, F.Sanger xc nh y cc trnh t phage X174 (5386 nucleotide vi 9 gene) v phage G4 (5577 cp nucleotide vi 10 gene), th n nay ngi ta xc nh v lp bn cho cc DNA c kch thc ln hn nhiu; v d: b gene phage lambda gm 48.502 cp base vi khong 61 gene (1983), nhim sc th s 3 ca nm men Saccharomyces cerevisiae gm 370.000 cp base (1992) v.v... ng k l, D n B gene Ngi (Human Genome Project = HGP; hnh 10.8) chnh thc i vo hot ng t 1990 do James Watson ch tr cng vi s cng tc ca nhiu nh khoa hc trn th gii. Vic phn tch trnh t b gene ngi c hon thnh vo 4/2003, cho thy b

gene (genome) ca chng ta c trnh t y gm 3.164.700.000 cp base, cha ng khong 25.000 gene m ha protein chu trch nhim xy dng nn mt b protein (proteome) gm khong 50.000 protein khc nhau trong cc t bo. HGP thc s l mt trong nhng k cng thm him v i nht trong lch s; ln u tin HGP cho php chng ta c c ton b thng tin di truyn ca t nhin lm nn con ngi (NHGRI 2005).
Hnh 10.8 Biu tng ca HGP cho thy tc ng ca d n ny ln nhiu lnh vc quan trng ca kinh t - x hi, cc vn v mi sinh, sc kho v i sng con ngi trn phm vi ton cu.
2. Cng ngh DNA ti t hp vi y-dc hc 2.1. Sn xut cc ch phm y-sinh hc bng cng ngh DNA ti t hp Nu nh vo nm 1972, Gio s Roger Guillemin (France) phi dng ti 500.000 no cu mi tinh chit c vi miligram somatostatin, mt hormone sinh trng ca vng di i th (vi cng trnh ny ng c trao gii Nobel nm 1980), th vo 10/1977 Hebert Boyer (USA) ch to thnh cng hormone ny t E. coli bng phng php DNA ti t hp. n 8/1978 chnh Boyer li l ngi u tin cho sn xut thnh cng insulin ngi t E. coli (hnh 10.9). Cc thnh tu u tin ny m ra mt k nguyn mi pht trin rc r nht trong lch s cng ngh sinh hc, cng ngh DNA ti t hp. Sau l hng lot cc ch phm khc nh cc hormone tng trng ca ngi (human growth hormone = HGH), b (BGH), ln (PGH), cc vaccine v cc interferon phng chng cc cn bnh nan ngi y nh ung th, vim gan v.v. v mt s bnh quan trng gia sc nh hin tng 'l mm-long mng' do virus gy ra... tt c u c sn xut t E. coli. c bit l, s ra i ca cc hng, cc cng ty ln u t mnh m cho s pht trin ca k ngh ny. Nh vy gp phn gii quyt mt cch c hiu qu nhiu vn thc tin t ra trong y hc, trong chn nui-th y, v nng nghip ni chung ...

nc ta cng c mt s cng trnh nghin cu v cc vn ny, chng hn nh nghin cu s sai khc di truyn gene hormon sinh trng ca mt s ging g Vit Nam. Qua cho thy intron I ca gene hormon sinh trng cc ging g Ri, g Ma, g c, g H di hn kch thc d on ca gene hormone sinh trng g c Tanaka cng b nm 1992 (Trn Xun Hon 2004).
Chuyn gene insulin enzyme gii hn cDNA ngi on ni Plasmid ti t hp
To dng gene insulin vi khun
chuyn gene
insulin insulin
insulin insulin
(a) (b) Hnh 10.9 (a) H. Boyer (tri) v S. Cohen; v (b) s th nghim to dng v biu hin gene insulin ngi vi khun E. coli.
Bn cnh vic sn xut cc hormone, vaccine... ni trn, ngi ta cn sn xut c cc khng th n dng (monocloning antibodies) dng : (i) xc nh vi khun gy bnh (nh thng hn chng hn); (ii) xc nh mc hormone t nh gi chc nng tuyn ni tit hoc s thay i trong qu trnh tng hp hormone do khi u gy ra; (iii) pht hin mt s protein c ngha trong chn on khi u hoc mt s tnh trng trc khi sinh; (iv) pht hin cc thuc b cm c trong mu, hoc kim tra nng thuc trong mu v t chc nhm m bo liu thuc s dng sao cho khng vt qu ngng gy c v.v. Nh c tnh c hiu v chnh xc cao v s dng d dng, vic sn xut cc khng th n dng to nn mt nhnh pht trin mau l nht ca cng ngh sinh hc, v cng vi vic sn xut cc vaccine chng tr thnh nhng phng tin quan trng nht trong chnh sch y t cng ng ca cc nc ang pht trin v xt v lu di, vic ng dng cc khng th n dng c trin vng cha c nhiu bnh trong c cc khi u c tnh. 2.2. ng dng k thut di truyn trong chn on v iu tr gene Trong chn on bnh mc phn t, thnh tu ni bt nht l vo nm 1981, ln u tin bnh thiu mu hng cu hnh lim c chn

on trc sinh mc gene nh phn tch DNA bng enzyme gii hn. T y m ra hai hng chnh trong chn on gene l: chn on cc bt thng bm sinh v chn on cc bt thng DNA soma. Mt s thnh tu khc t c theo hng u gm c: bnh hemophilia A, ri lon dng c Duchenne, hi chng X-fragile, bnh retinoblastoma - mt dng ung th vng mc...Theo hng sau, ngi ta p dng i vi mt s dng ung th mu nh cc lympho Burkitt, lympho nang, bnh bch cu...ng k l gn y, ngi ta xc nh c nhiu gene gy bnh quan trng ngi, nh: gene gy bnh ha x nang (cystic fibrosis) nm 1990, gene gy bnh Huntington nm 1993... Liu php gene (gene therapy) cng l mt phng thc sn xut v iu tr mi bng cc phn t tr liu. y l hng c nhiu trin vng nht nhng kh thc hin nht v n c lin quan n c vn phng php lun ln kha cnh o l. S kin ni bt nht l vo nm 1990-91, W.F.Anderson, R.M.Blaese v Ken Cuver thc hin thnh cng ca liu php gene u tin trn mt b gi bn tui mc bnh suy gim min dch phi hp (SCID) vi s thiu ht adenosine deaminase (ADA), mt enzyme cn thit cho sn xut cc khng th trong cc t bo h mim dch, gi l hi chng "bubble-boy" (bnh do gene gn u mt vai ngn nhim sc th s 20 gy ra). Mt khc, liu php gene cng m ra nhng trin vng to ln trong cha tr cc cn bnh ph bin nht, tc ng ti hng triu triu ngi trn hnh tinh nh: ung th, SIDA/AIDS, vim gan do virus, cc bnh tim mch, cc bnh thoi ha thn kinh (bnh Parkinson, bnh Huntington, bnh Alzheimer...) hay c nhng bnh mn tnh nh a thp khp. 2.3. K thut di truyn vi hnh php hc v mt s vn x hi khc K thut di truyn cn c ng dng rt hiu qu trong cc ngnh hnh php hc, chng hn bng cch s dng k thut 'du vn DNA' (DNA fingerprinting) thay cho 'du vn tay' trc y cho php cc ngnh cnh st hnh s xc nh chnh xc cc ti phm hoc nhn dng xc nn nhn trong chin tranh hoc do tai nn gy ra (hnh 10.10).
Hnh 10.10 Du vn DNA (DNA fingerprinting) c th gip cc nh nghin cu xc nh kh nghi trong trng hp ti phm. Kiu vch ngang biu th bn cht di truyn ca mt ngi. mu ny cho thy cc bng ca mu ngi mang m s S2 kh nghi trng khp vi bng chng, mu mu E(vs).

nc ta, trong thi gian gn y, k thut ny v ang c p dng mt cch hiu qu trong ngnh hnh s. Bn cnh c cng trnh phn tch DNA nhn din t bo trn 103 ngi Vit Kinh bng k thut PCR - SSO (Kit Innolipa). Qua phn tch so snh khong cch gene hc gia ngi Vit Kinh v 11 sc tc chu v i dng khc, kt lun rng ngi Vit Kinh gn vi ngi Thi ri n ngi Manchu (V Triu An, 1999). 3. K thut di truyn vi cc sinh vt bin i gene (genetically modified organisms = GMOs) i vi ngnh chn nui, cng ngh sinh hc ni chung v cng ngh sinh hc ni ring t nhiu thnh tu ng k, chng hn cc k thut chuyn ghp gene p dng cho hp t v phi cc gia sc nhm tng cng kh nng chng bnh v ci thin ging ni chung; cng nh cc k thut mi trong xc nh gii tnh ca phi... Hnh 10.11 cho thy kh nng ng dng k thut cy ghp gene trn trng chut c th tinh. Sau em phi ghp gene cy vo trong t cung ca con chut lm m khc. Kt qu l to ra c chut con c b lng dng khm nh mong mun. in hnh cho cc th nghim truyn gene ng vt l vo nm 1982,R.D.Palmiter, R.L.Brinster v cc ng s i hc Seatle (Philadelphia, USA) bng cch truyn gene xc nh hormone sinh trng ca chut cng vo trng th tnh ca chut bnh thng, ri cy tr li cc t bo
Tinh chit gene Tim
cc trng chut
Phi c cy trong t cung ca chut m thay th
i con
Hnh 10.11 M hnh tng qut v th nghim truyn gene ng vt (xem gii thch trong bi).
c bin i gene vo vi trng ca cc chut ci c th mang thai cho n cng. V kt qu l, cc tc gi ny thu c dng chut nht c kch thc ln gp 2-3 ln chut bnh thng, gi l chut khng l. i vi trng trt, vic s dng cc phng php chuyn ghp gene t c nhiu thnh tu to ln. Chng hn, hng Biogen (USA) nm

1984 chuyn thnh cng plasmid Ti vo t bo thc vt; hng Calgene v Phytogene (USA, 1984) ghp thnh cng gene khng glyphosate bo v cy bng; nm 1985 hng Molecular Genetics (USA) to c ging ng mi cho nhiu tryptophan. Nm 1993, bng k thut sng bn gene vo t bo thc vt ngi ta a c gene sn xut protein dit su vo cy ng, v kt qu l to ra c ging ng chng chu cao i vi su c thn. iu th v l vic tch cc gene c nh m, gene Nif (Nif = nitrogene fixation) t cc vi khun nt sn cy h u v a vo b gene ca cc cy trng khc to ra cc ging cy trng mi c kh nng c nh nit v cho nng sut cao. Trong chn ging vi sinh vt, ngi ta thc hin thnh cng vic chuyn gene cellulase vo vi khun (J.P.Aubert, France 1/1983), ci bin E. coli sn xut L-aspartat (hng Tanabe, Japan 1985), ghp gene vo x khun S. violacconiger ci tin vic sn sinh enzyme glucoisomerase (hng Roquette v Cayla, France 1985), ngoi ra cn to c cc ging vi sinh vt bin i gene c kh nng n cn du dng trong x l cc ph thi c c t nhm bo v mi sinh. Bn cnh vic to ra ging nm men mi c th git cht cc vi khun xut hin trong bia (hng Suntory, 1985), cn to c chng nm men sn xut insulin v interferon (A. Kimura, Japan 1986) v.v. Nhn thc r ngha v tm quan trng ca cng ngh DNA ti t hp v cng ngh sinh hc ni chung i vi s pht trin kinh t-x hi t nc ta trong th k XXI, Chnh ph ra Ngh quyt 18/CP ngy 11/4/1994 v "Phng hng pht trin ca cng ngh sinh hc Vit Nam n nm 2010". y c xem l mt bc ngot quan trng cho s pht trin ca cng ngh sinh hc nc nh trong thi gian qua v sp ti.
Cu hi v Bi tp
1. Th no l enzyme gii hn? Chng c nhng tnh cht no m c coi l cng c thit yu i vi cng ngh DNA ti t hp? Bng hai v d v enzyme gii hn, hy chng t rng t nht c hai phng php thnh lp cc phn t DNA ti t hp in vitro. 2. T cc enzyme gii hn BamHI, EcoRI v HindIII Bng 10.1 v BglII c on ch c bit l AGATCT, hy cho bit cp enzyme no s to ra cc u dnh hay u b sung (sticky/complementary ends) tng thch? Trnh t ca u dnh tng thch ny l g? 3. Gi s bn ct hai DNA khc nhau, mt vi BamHI v mt vi BglII, sau ni chng li vi nhau thng qua cc u dnh tng thch.

Mt khi khu ni ri, bn c th tch hai DNA ny ln na bng mt trong hai enzyme gii hn hay khng? Ti sao, hoc ti sao khng? 4. Gi s bn bin np mt DNA ti t hp cho mt vi khun v do nhm ln, bn t cc t bo c bin np trn mi trng khng c cht khng sinh no c. Kt qu m bn quan st c l g? Ti sao? 5. Gi s rng bn chit xut c mt enzyme ct gii hn t loi vi khun c tn khoa hc l Xenobacterium giganticus. Theo danh php hc, bn s vit tn enzyme nh th no? 6. Cho bit trnh t ca mt on DNA c cha mt v tr nhn bit i xng xui ngc (palindromic recognition site) cho mt enzyme gii hn l GACGATATCAACT. Hy tm trnh t ca v tr nhn bit . 7. Th no l phn t DNA ti t hp, vector tch dng? Hy nu cc bc ca quy trnh to dng gene ti t hp v cho s minh ho. 8. Gi s tinh ch c plasmid pBR322 v phn t DNA ngi c cha mt gene cn nghin cu. Hy phn tch k thut to dng gene ni trn E. coli. 9. Enzyme phin m ngc l g? N c tinh ch t loi sinh vt no v c ng dng vo khu no trong cng ngh DNA ti t hp? Gii thch v cho s minh ho. 10. C th s dng cc cht khng sinh xc nh xem liu plasmid pBR322 nhn c mt on xen trong v tr EcoRI ca n hay khng? Ti sao, hoc ti sao khng?
Ti liu Tham kho

Ting Vit Trn Xun Hon. 2004. Nghin cu s sai khc di truyn gene hormone sinh trng ca mt s ging g Vit Nam. Lun n Tin s Di truyn hc, Th vin Quc gia, H Ni. L nh Lng. 2001. Nguyn l K thut Di truyn. NXB Khoa hc v K thut, H Ni. Phan C Nhn. 1999. Cng ngh DNA ti t hp. Trong: Di truyn hc tp II (Phan C Nhn, ch bin). Trang: 257-303. NXB Gio Dc, H Ni. Hong Trng Phn. 1995. Mt s vn v Di truyn hc hin i (Ti liu BDTX cho gio vin THPT chu k 1993-1996). Trng HSP Hu. Hong Trng Phn. 1997. Di truyn hc Phn t. NXB Gio Dc. Hong Trng Phn. 1999. "Gii thiu cng ngh sinh hc"; v "C s

khoa hc ca cng ngh sinh hc". Trong: Chuyn Cng ngh Sinh hc (Ti liu BDTX gio vin THPT chu k 1997-2000; bin son chung vi Nguyn B Lc v Bin Vn Minh). Trang: 56-96. Trng HSP Hu.
Ting Anh
Vu Trieu An 1999. Mitochondrial DNA polymorphism in the Vietnamese population. Eur. J. of Immunogenetics, 26: 471-422. Campbell NA, Reece JB. 2001. Essential Biology. Benjamin/Cummings, an imprint of Addison Wesley Longman, Inc, San Francisco, CA. Collins FS, Green ED, Guttmacher AE DNA Guyer MS. 2003. A Vision for the Future of Genomics Research. Nature, Vol.422, No.6934, p.835-847. Hartwell, Hood, Goldberg, Reynolds, Silver, Veres. 2004. Genetics From Genes to Genomes. 2nd Edition. McGraw Hill, Inc., New York. Lewis R. 2003. Human Genetics: Concepts DNA Applications. 5th ed, McGraw-Hill, Inc, NY. Palladino MA. 2002. Understanding the Human Genome Project. Benjamin/Cummings Publishing Company, Inc, Menlo Park, CA. Watson JD, Hopkins NH, Roberts JW, Steitz JA, Weiner AM. 1987. Molecular Biology of the Gene. 4th ed, Benjamin/Cummings Publishing Company, Inc, Menlo Park, CA. Watson JD, Gilman M, Witkowski J, Zoller M. 1992. Recombinant DNA. 2nd ed, Scientific American Books, New York. Weaver RF, Hedrick PW. 1997. Genetics. 3rd ed, McGraw-Hill Companies, Inc. Wm.C.Browm Publishers, Dubuque, IA. Mt s trang web Agricultural Biotechnology http://www.aphis.usda_gov/biotechnology/ http://www.agbioworld.org Online Mendelian Inheritance in Man (OMIMTM): http://www.ncbi.nlm.nih.gov/Omim/mimstats.html http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=OMIM National Human Genome Research Institute (NHGRI): http://www.ncbi.nlm.nih.gov/genome/guide/human The Institute for Genomic Research: http://www.tigr.org Celera Genomics Corp.: http://www.celera.com

Chng 11
Di truyn hc Ngi
I. Cc phng php nghin cu di truyn hc ngi
1. Phng php phn tch ph h (Genealogy analysis) Phng php phn tch ph h c s dng nghin cu s di truyn cc tnh trng ngi thuc cng dng h, xc nh c tnh trng hoc bnh no l tri hay ln..., do mt hay nhiu gene qui nh, c tnh cht di truyn hay khng, di truyn c lp hay lin kt vi gii tnh..., kh nng mc bnh ca cc th h tip theo. Trong mt s trng hp cn xc nh c ngi d hp t mang gene bnh. Phng php ny kt hp vi cc xt nghim khc cho php c th rt ra nhng li khuyn v di truyn chnh xc v hu ch cho cc gia nh v vic sinh con hoc kt hn. Phng php ny c p dng khi bit c cc t tin trc tip v con chu ca ngi bnh qua nhiu th h. Mt s k hiu dng lp ph h Nam bnh thng Nam bnh N bnh thng Gii tnh cha c xc nh N bnh Cp sinh i khc trng
Cp v chng Cht
Cp sinh i cng trng

Th d hp mang gene bnh trn nhim sc th thng
Th d hp mang gene bnh lin kt vi nhim sc th X Th h I Hn nhn

Cc con lit k t tri sang phi theo trnh t
II
1 2 3 4 5
III

2. Phng php nghin cu tr sinh i C hai loi sinh i l sinh i cng trng (monozygotic twin-MZ) v sinh i khc trng (dizygotic twin-DZ). Ngi ta da vo hng lot c im v s lng v cht lng phn bit tr sinh i cng hay khc trng: tr sinh i cng trng bt buc cng gii, tr sinh i khc trng c th cng hay khc gii; tr sinh cng trng c mt mng m chung, tr sinh khc trng c mng m khc nhau; tr sinh cng trng khi ghp m th lun lun thnh cng; c s ging nhau tr sinh i cng trng v s khc nhau tr sinh i khc trng v nhiu tnh trng. Thng thng, chn nhng tnh trng di truyn r rt, t b bin i di nh hng ca cc nhn t mi trng, thuc nhng tnh trng ny c nhm mu, sc t mt, da v tc, np vn tay, chn. Trong phn ng cy ghp m l phng php c th kt lun mt cch chnh xc nht. So snh cc cp sinh i v mt tnh trng hoc mt bnh no cho php nh gi nh hng ca yu t di truyn v yu t mi trng ln s hnh thnh tnh trng, pht hin cc bin d xy ra do yu t mi trng... Mt bnh hoc mt tnh trng di truyn no c th biu hin c hai thnh vin ca cp sinh i (c tng hp) hoc cng c khi ch biu hin mt trong hai thnh vin ca cp sinh i (khng tng hp). cc cp sinh i cng trng nu tng hp cng ln th vai tr ca yu t di truyn cng ln. Nu tng hp cng nh th vai tr ca yu t di truyn cng km. 3. Phng php di truyn t bo hc ngi y l phng php c dng ph bin hin nay pht hin v quan st nhim sc th, qua xc nh cc d dng nhim sc th, cc hin tng lch bi, hin tng cu trc li nhim sc th dn n nhiu bnh di truyn him ngho ngi. Dng m gm nhiu t bo ang phn chia mnh nh m tu xng , m bo thai, m tinh hon, khi u c tnh...lm tiu bn phn tch, nh gi nhim sc th. K thut lai t bo soma v k thut hin bng nhim sc th ra i cho php nghin cu c ch ung th, hot ng ca gene trong qu trnh pht trin c th v gp phn tch cc vo vic lp bn di truyn ngi. 4. Phng php nghin cu qun th Phng php ny da vo phng trnh Hardy - Weinberg, nh gi tn s cc kiu hnh tnh mt cc gene trong qun th lin quan n cc bnh di truyn. N cn cho php nh gi cc hu qu ca giao phi cn huyt v theo di s di truyn ca cc qun th ngi v mt ngun

gc. 5. Cc k thut sinh hc phn t Cc thnh tu to ln trong nhng nm gn y v lnh vc Di truyn hc ngi, c bit l thnh tu gii m b gene ngi t c l nh s dng cc k thut sinh hc phn t nh tch chit, phn tch nh tnh v nh lng nucleic acid; cc phng php lai phn t: Southern blot, Northern blot, lai ti ch (in situ hybridization),... ; cc phng php xc nh trnh t nucleic acid; to dng (cloning); xy dng th vin b gene, th vin cDNA; phng php PCR (polymerase chain reaction); Sinh tin (Bioinfomatics),... Ngoi cc phng php trn ngi ta cn s dng cc phng php nghin cu m phng hc, phng php in v phn tch np vn da, phng php iu tra dch t hc v phng php di truyn lm sng... trong nghin cu Di truyn hc ngi.
II. Cc phng php lp bn di truyn ngi

1. Phn tch lin kt (Linkage analysis) S trao i cho gia cc locus trn cng nhim sc th c th to ra ti t hp. Vic nh gi tn s ti t hp c th thc hin da vo quan st s di truyn cc allele trong cc gia nh. S xc nh cc phase lin kt (c ngha l nhim sc th m trn mi mt allele c nh v) l mt phn quan trng ca phng php ny. Mc d c s tng quan gia centiMorgan v khong cch vt l tht s gia cc locus, mi quan h ny b phc tp do cc sai khc v gii tnh trong ti t hp, cc tn s ti t hp cao hn gn cc telomere v s tn ti ca cc im nng ti t hp (recombination hot spots). S sai khc v thng k ca 2 locus c th c tnh ton bng cch tnh t s ca hai hp l (likelihood): hp l ca lin kt tn s ti t hp cho chia cho hp l ca khng lin kt. Logarithm ca t s sai khc ny c k hiu l LOD (logarithm of odds). LOD>0,3: c lin kt; LOD<0,2: khng lin kt. Khong cch gia cc locus c tnh bng n v centiMorgan (cM). 1 cM tng ng vi tn s ti t hp xp x 1%. Phn tch lin kt cho php chng ta xc nh c khong cch tng i gia cc locus nhng khng xc nh c cc v tr c trng vi marker hoc cc gene bnh. 2. Cc phng php lp bn vt l (physical mapping) 2.1. Phng php lp bn mt on (delection mapping)

Karyotype ca cc bnh nhn mc bnh di truyn thnh thong c pht hin thy hin tng mt on mt vng c trng trn nhim sc th. iu ny cho thy rng locus gy bnh c th nm trong vng b mt. Chiu di on b mt c th bin i mt s bnh nhn mc cng mt bnh. Cc on mt ca nhiu bnh nhn c so snh xc nh vng b mt tt c cc bnh nhn, do thu hp li v tr ca gene bnh. Phng php lp bn mt on c s dng xc nh v tr cc gene chu trch nhim i vi ung th vng mc (retinoplastoma), hi chng Prader Willi v Angelman v khi u Wilms. Khi u Wilms l mt khi u thn giai on u do t bin nhim sc th s 11 gy ra. Ch rng cc mt on ny ch xy ra mt nhim sc th trong cp tng ng, to ra bnh nhn d hp t i vi on b mt. Nu on b mt ln c th quan st d dng di knh hin vi v xy ra trn c hai nhim sc th ca cp tng ng th thng gy cht. 2. 2. Phng php lai ti ch (In situ hybridization) on DNA thu c t marker a hnh hoc gene bnh m chng ta mun bit v tr c to dng vi mu d (probe) bng k thut DNA ti t hp. Mu d c nh du vi cht phng x nh tritium chng hn. Sau mu d c t vo mt mng lai cha nhim sc th k gia m DNA ca n bin tnh. Mu d phng x s bt cp b sung vi DNA bin tnh ca on nhim sc th c trng. Bi v mu d pht ra bc x nn v tr ca n c th c xc nh mt cch chnh xc bng cch t mt phim nhy cm tia X ln trn mng lai (phng x t ghi (autoradiography)). Phng php lai ti ch s dng cc mu d phng x thng din ra chm v mc phn tch kh thp (2-5 Mb). K thut FISH (fluorescence in situ hybridization) s dng mu d hunh quang thay cho mu d phng x l nhanh hn v cho php phn tch vng ca mu d tt hn, thng l trong khong 1 Mb i vi nhim sc th k gia. Bng cch s dng nhim sc th k trung gian, cc nhim sc th k ny kt xon t hn cc nhim sc th k gia, phn tch FISH c th c tng ln t 25-250 kb. 2.3. Lai t bo soma Cc t bo soma ca cc loi khc nhau khi sinh trng trong cng mi trng nui cy vi s c mt ca mt s tc nhn nh polyethylene glycol hoc virus Sendai, thnh thong s dung hp vi nhau to thnh cc t bo lai. Khi lai cc t bo chut v t bo ngi vi nhau s thu c cc t bo lai cha 86 nhim sc th : 46 nhim sc th ngi v 40 nhim sc th chut. Sau cc t bo lai c php sao chp. Cc t bo ny bt u mt i mt s nhim sc th ngi khi chng tri qua qu trnh nguyn

phn. Cui cng cc t bo cn li b nhim sc th y ca chut v ch mt hoc mt s nhim sc th ngi. Cc t bo ny c lp karyotype xc nh nhim sc th no ca ngi cn li (nhim sc th ca ngi v chut c th c phn bit vi nhau da vo kch thc v cc phng php hin bng). Sau cc t bo ny c nghin cu xc nh t bo no mang gene thch hp ang ni n. V d nu gene lun lun c tm thy trong cc t bo cha nhim sc th s 1 ca ngi nhng cha bao gi c pht hin trong cc t bo khng mang nhim sc th s 1, chng ta c th kt lun rng gene ny phi c nh v trn nhim sc th s 1. S c mt ca gene trong t bo lai c th c pht hin bng mt s phng php. Nu gene m ho mt enzyme c sn xut ra bi t bo th mt xt nghim enzyme c th c s dng. in di protein c s dng nhn ra mt sn phm protein ngi v phn bit n vi sn phm protein tng ng ca chut. Tuy nhin cc phng php ny yu cu mt sn phm protein bit phi c pht hin. Thng thng hn, hin nay cc nh nghin cu kim tra s lai ca cc dng t bo dung hp vi mt mu d nh du phng x mang DNA quan tm bng cch s dng phng php Southern blotting hoc k thut PCR. 2.4. Lp bn lai phng x (Radiation hybrid mapping) Phng php ny bt u vi mt th lai t bo soma cha mt nhim sc th n ca ngi. Sau cc t bo ny c chiu vi tia X hoc tia lm t gy si kp nhim sc th. Bc x ny git cht t bo v vy chng phi c dung hp vi t bo ng vt gm nhm tn ti. Mt s t bo lai c to ra gi l cc th lai phng x (radiation hybrid) mang cc on nh nhim sc th ngi lai vi nhim sc th gm nhm. S c mt ca nhim sc th ngi c th c pht hin bng s sng lc i vi cc trnh t Alu. Cc trnh t Alu ny tm thy mi mt on c kch thc vi kb trong nhim sc th ngi nhng khng tm thy trong nhim sc th ca gm nhm. Bi v bc x b gy nhim sc th ngi nhng khong cch ngu nhin, cc locus c nh v gn vi mt locus khc hn s c tm thy thng hn trn cng on nhim sc th (c xu hng cng i vi nhau). K thut PCR c th c s dng xc nh th lai phng x mang cc t hp c trng ca cc locus ngi. Sau s dng phng php thng k nh gi mi mt cp locus thng c tm thy trn cng on nhim sc th nh th no. Kt qu ny cung cp s nh gi khong cch tng i gia cc locus. Ngoi cc phng php trn ngi ta cn dng cc phng php to dng nh v (positional cloning), phn tch s bo tn ca cc loi lai

(analysis of cross-species conservation), phn tch trnh t DNA bng my tnh, sng lc cc t bin trong trnh t (screen for mutations in the sequence), kim tra s biu hin ca gene.... III. Nhim sc th Y v cht nhim sc gii tnh ca ngi 1. Nhim sc th Y ca ngi Tri ngc vi nhim sc th X, nhim sc th Y hon ton nh v cha rt t gene. Phn ln chiu di ca nhim sc th Y khng xy ra ti t hp l cht d nhim sc. Cc on lp DNA ca n c xem xt t m. Cc vng tng ng ca cp nhim sc th XY c nh v mi u ca nhim sc th Y. Cc vng ny kt cp v ti t hp vi nhim sc th X trong k u ca gim phn I. Chng c gi l cc vng nhim sc th thng gi (pseudoautosomal region-PAR) bi v bt k gene no nh v trong cc vng ny Hnh 11.1 (cho n nay ch mi pht hin c 9 gene) Nhim sc th u c di truyn hon ton ging nh gene Y ca ngi trn nhim sc th thng. Nam c hai bn sao ca cc gene ny: mt trong PAR ca Y v mt trong vng tng ng ca X. Khong 95% chiu di ca Y nm gia cc PAR to ra vng khng tip hp (nonrecombining region-NRY). Gn 80 gene c tm thy y. Chng c xp thnh 2 nhm chnh. Cc gene thuc nhm th nht (m ho cc protein s dng cho tt c cc t bo (c hai gii)) c bu hin nhiu m. Cc gene gi nh ny (housekeeping gene) c th tng ng trn X m trnh khi s bt hot X n. Cc gene thuc nhm th hai m ho cc Hnh 11.2 protein hot ng chc nng ch trong tinh S tip hp ca hon v khng c th tng ng trn X. Mt nhim sc th X gene c vai tr quan trng trong nhm ny l v Y SRY (sex-determing region Y). SRY nh v trn cnh ngn nm bn ngoi PAR v gy ra cc kh nng bin i phi thnh nam gii.

2. Cht nhim sc gii tnh ca ngi t lu ngi ta bit rng n c hai nhim sc th X trong khi nam ch c mt nhim sc th X. Nh vy mi gene lin kt vi nhim sc th X c hai bn sao i vi n v mt bn sao i vi nam. Nu tt c cc gene trn nhim sc th X u hot ng th n hm lng cc cc sn phm (nh enzym chng hn) do cc gene ny m ha phi nhiu gp i nam. Trong thc t hm lng cc sn phm trn u Hnh 11.3 S bt hot ca nhim sc th X bng nhau c hai gii. C th gii thch iu ny nh th no ? Vo u thp nin 1960, Mary Lyon gi thuyt rng mt nhim sc th X trong mi t bo soma ca n b bt hot. Hin tng ny dn n kt qu c gi l s b p v liu lng gene (Gene dosage compensation) lm cho sn phm ca gene lin kt vi nhim sc th X ca nam v n ngang nhau. Gi thuyt Lyon cho rng s bt hot nhim sc th X xy ra sm trong giai on pht trin phi ging ci. mt s t bo, nhim sc th X b bt hot c ngun gc t b, trong khi nhng t bo khc nhim sc th X b bt hot c ngun gc t m. Qu trnh bt hot ny l ngu nhin. Khi mt nhim sc th X b bt hot trong mt t bo, n s duy tr s bt hot ny tt c cc th h t bo con chu. Nh vy s bt hot nhim sc th X l ngu nhin nhng li c c nh. S bt hot nhim sc th X s dn n kt qu l tt c c th ci bnh thng c hai qun th t bo phn bit: mt qun th c nhim sc th X hot ng nhn c t b v mt qun th c nhim sc th X hot ng nhn c t m. Vi hai qun th t bo ny, ging ci l dng khm i vi nhim sc th X, ging c ch c mt bn sao ca nhim sc th X l dng bn hp t (hemizygous) i vi nhim sc th X. Gi thuyt Lyon a ra mt s bng chng, phn ln nhn c t cc nghin cu v ng vt v ngi. Cc nghin cu ca Di truyn hc trong thp k 1940 cho thy rng cc t bo gian k ca mo ci thng cha mt khi cht nhim sc dy c bt mu thuc nhum trong nhn t bo. Cc khi ny cha bao gi tm thy ging c. Chng c gi l th Barr, do Murray Barr, mt nh Di truyn hc m t chng. Barr gii thch rng th Barr tng

ng vi mt nhim sc th X kt c cao. Trng thi kt c ny phn nh thc t l ADN ca n c ti bn chm hn ADN cc nhim sc th khc trong pha S. Cc nghin cu tip theo xc nh mARN c sao chp ch t mt nhim sc th X trong t bo ca con ci bnh thng. Qu trnh bt hot xy ra khong hai tun sau khi th tinh. Qu trnh c khi u mt v tr n (single location) trn cnh di ca nhim sc th X (trung tm bt hot X) v sau tri di dc theo nhim sc th. S bt hot ca nhim sc th X l thng xuyn i vi tt c cc t bo soma con ci, nhng n phi tr nn hot ha trong dng t bo mm ca c th ci, do vy mi t bo trng s nhn c mt bn sao ca nhim sc th X hot ng. Theo gi thuyt Lyon, s lng th Barr trong cc t bo soma l lun lun bng s nhim sc th X tr i 1. C th ci bnh thng c mt th Barr trong t bo soma trong khi cc con c li khng c. Nam b hi chng Klinefelter (XXY) c mt th Bar. Dng hi chng ny dn n mt cu hi: Nu nhim sc th X tha b bt hot th ti sao ngi mang nhim sc th X tha c kiu hnh khng bnh thng ? Cu tr li l s bt hot ca nhim sc th X l khng hon ton. Mt vi vng ca nhim sc th X bt hot vn hot ng tt c cc bn sao. Mt nghin cu mi y cho thy 20% gene ca nhim sc th X bt hot c th vn hot ng. Cc gene ny l tng ng vi cc gene trn nhim sc th Y v cc bn sao ca chng gp phn to nn s khng bnh thng ca kiu hnh. Trung tm bt hot X cha gene cn thit cho s bt hot c tn l XIST (X-inactive specific transcript), gene ny c sao m thnh mARN 15-17 kb nhng khng c dch m. Mt im c bit khc ca nhim sc th X bt hot l s methyl ha, c kh nng l s methyl ha chu trch nhim duy tr s bt hot ca mt nhim sc th X c bit trong t bo.
IV. S di truyn cc gene tri ln trn nhim sc th thng v nhim sc th gii tnh
1. S di truyn cc gene tri ln trn nhim sc th thng 1.1. S di truyn cc gene tri trn nhim sc th thng Bnh di truyn do cc gene tri trn nhim sc th thng c bt gp vi tn s khong 1/200. Nu tnh ring l th mi bnh di truyn do gene tri trn nhim sc th thng l kh him trong qun th. Tuy nhin, cc bnh ph bin nht c tn s gene khong 0,001. Tnh trng hay bnh di truyn do cc gene tri trn nhim sc th thng c mt s cc c

im quan trng. Th nht, c hai gii biu hin tnh trng vi t l gn nh nhau. B v m c vai tr ngang nhau trong vic truyn tnh trng cho con ci. Th hai, khng c s ngt qung th h: kiu hnh bnh c truyn t ng b sang b m, ri t b m cho con ci...Nu b m khng b bnh th s khng truyn bnh cho con ci. V vy kiu biu hin ny c gi l kiu di truyn phn b dc (vertical transmission pattern). Th ba, mc d s truyn tnh trng t b cho con trai khng c yu cu nh gi s di truyn gene tri trn nhim sc th thng nhng s hin din ca n trong mt ph h loi tr chc chn cc kiu di truyn khc (c bit l s di truyn lin kt vi nhim sc th X). Cui cng, th d hp t mang gene bnh truyn tnh trng ny cho gn mt na con ci ca h. 1.2. S di truyn cc gene ln trn nhim sc th thng Ging nh bnh di truyn do cc gene tri trn nhim sc th thng, cc bnh di truyn do gene ln trn nhim sc th thng l kh him trong qun th. Ch khi trng thi ng hp th mi biu hin thnh kiu hnh. Cc th d hp l ph bin nhiu hn th ng hp. S di truyn cc gene ln trn nhim sc th thng c mt s c im quan trng. Th nht, kiu hnh bnh biu hin mt hoc nhiu anh ch em nhng khng pht hin thy cc th h trc . Do vy kiu di truyn ny c gi l kiu di truyn phn b ngang (horiziontal transmission pattern). Th hai, ging nh trng hp di truyn do gene tri trn nhim sc th thng, nam v n u c kh nng b mc bnh vi t l ngang nhau. Th ba, trung bnh khong 1/4 con ci ca cp b m d hp s b mc bnh. Cui cng, tnh ng huyt (consanguinity) thng hin din trong cc ph h lin quan n bnh do gene ln trn nhim sc th thng nhiu hn so vi cc ph h lin quan n cc kiu di truyn khc. 2. S di truyn cc gene tri ln trn nhim sc th gii tnh 2.1. S di truyn gene ln lin kt vi nhim sc th X Mt s cc tnh trng v bnh ni ting l do cc gene ln lin kt vi nhim sc th X gy ra. Cc kiu di truyn do gene ln lin kt vi nhim sc th X chc chn l khc vi gene trn NST thng. Bi v n c di truyn hai bn sao nhim sc th X. n, tnh trng do gene ln lin kt vi nhim sc th X rt ging vi trng hp tnh trng do gene ln trn nhim sc th thng quy nh. Tuy nhin, ch c mt nhim sc th X l bt hot trong t bo, nh vy mt na s t bo ca n d hp t s biu hin allele bnh v na cn li s biu hin allele bnh thng. Trong lc nam c di truyn mt allele bnh ln lin kt vi nhim sc th X th s biu hin bnh v nhim sc th Y khng mang allele tng ng.

Mt bnh do gene ln lin kt vi nhim sc th X vi tn s q th tt c nam c t bin s biu hin bnh. N phi cn hai bn sao ca allele t bin biu hin bnh do tn s biu hin s l q2 nh trng hp bnh do gene ln trn nhim sc th thng quy nh. Kt qu ny cho thy rng nam b tc ng mt cch thng xuyn v nhiu hn so vi n i vi bnh do gene ln lin kt vi nhim sc th X. Mt s c im phn bit tnh trng hoc bnh do ln lin kt vi nhim sc th X vi tnh trng hoc bnh do gene ln trn nhim sc th thng: - Cc gene lin kt vi nhim sc th X khng c truyn trc tip t b cho con trai. - Gene lin kt vi nhim sc th X c th c truyn t b cho tt c con gi, nhng con gi ny khng c biu hin bnh nhng c mang gene bnh. Cc con gi mang gene bnh ny s truyn li cho mt na s con trai ca h, cc con trai ny b bnh. Kiu di truyn ny gy ra s xut hin cc th h ngt qung (skipped generations). Kiu ghp i ph bin i vi cc gene ln lin kt vi nhim sc th gii tnh X l s kt hp ca m mang gene bnh vi b bnh thng. M s truyn gene bnh cho mt na s con trai v mt na s con gi. Mt kiu ghp i ph bin khc l gia b b bnh vi m bnh thng. Tt c con trai ca h bnh thng. Bi v tt c con gi phi nhn nhim sc th X ca b do chng s tr thnh th mang gene bnh d hp t. Nh vy khng c mt a con no biu hin bnh. Kiu ghp i t ph bin hn l gia b b bnh vi m l th mang gene bnh. Mt na s con gi ca h s tr thnh th mang gene bnh d hp t, mt na s ng hp t gene bnh v s biu hin bnh. Mt na s con trai bnh thng v mt na s con trai b bnh. Thnh thong c trng hp n ch nhn mt bn sao n ca gene ln gy bnh lin kt vi nhim sc th gii tnh m c th b bnh. Cc c th n ny c gi l th d hp biu hin (manifesting heterozygotes). Bi v cc c th n ny thng duy tr t nht mt phn nh ca nhim sc th X hot ng bnh thng v c khuynh hng biu hin bnh tng i nh. V d, gn 5% n d hp t i vi bnh a chy mu A (hemophilia A) c mc yu t ng mu VIII thp c xp vo loi bnh a chy mu nh. t ph bin hn l n ch c mt nhim sc th X (hi chng Turner) biu hin cc bnh do gene ln lin kt vi nhim sc th X, nh bnh a chy mu A. N cn c th b cc bnh do gene ln lin kt nhim sc

th X l kt qu ca s chuyn on hoc mt on nhim sc th X. Cc trng hp ny l him. 2. 2. S di truyn cc gene tri lin kt vi nhim sc th X S di truyn do gene tri lin kt vi nhim sc th gii tnh X ging nh s di truyn do gene tri trn nhim sc th thng. Mt c th ch cn tha hng mt bn sao n ca gene tri lin kt vi nhim sc th X l c th biu hin tnh trng hoc bnh. Bi v n c hai nhim sc th X, m mi nhim sc th X u c th c kh nng mang gene bnh nn n b bnh gn gp i so vi nam (tr trng hp ri lan gy cht nam). B b bnh khng th truyn tnh trng ny cho con trai. Tt c con gi u di truyn gene bnh ny v th u b bnh. B b bnh thng l d hp t v vy s c 50% c hi truyn allele bnh cho con gi v con trai. 2.3. S di truyn lin kt vi nhim sc th Y S tnh trng hoc bnh bit do gene trn nhim sc th Y qui nh l rt t, ch gp nam gii v c truyn trc tip t b cho con trai mang tnh cht dng h ni. Tt c cc con trai u b bnh (di truyn thng) v con gi khng mang bnh. Cc bnh lin kt vi nhim sc th Y nh tt nhiu lng mc vnh tai, dy sng lng bn tay, tt dnh ngn s 2 v 3...
V. Di truyn y hc
1. Cc bnh di truyn do ri lon chuyn ho v cc bnh nhim sc th 1.1. Cc bnh di truyn do ri lon chuyn ho 1.1.1. Phenylketonuria (PKU) PKU l mt d tt bm sinh, c di truyn do gene ln Mendel. PKU c Phelling pht hin ln u tin vo vo nm 1934. Tr s sinh b bnh ny l do t bin gene, gan khng c kh nng tng hp phenylalanine hydroxylase nn dn n tnh trng khng ch km hm s chuyn ho phenylalanine thnh tyrozine m cn gy ra ng phenylalanine trong mu, lm tng s phn gii phenyalanine thnh axit phenylpyruvic, cng b tch t trong mu. C phenylalanine v phenylpyruvic khi ln no nhiu s u c t bo thn kinh. Bn thn nc tiu thi ra cng c phenylalanine. Nu t bin gene ln xy ra mt khu khc lm km hm hot ng ca enzyme tyrozinase, l enzyme xc tc phn ng chuyn ho tyrozine thnh melanine dn ti bch tng. Nhiu nc ngy nay t ra th tc chn on cho mi tr s sinh, pht hin sm dng ng hp PKU cho n khu phn king c

bit ngho phenylalanine (cho sa nga) hoc khu phn n khng c phenylalanine (rau, mt ong, b,...) 1.1.2. Alcaptonuria Bateson,1902 v Garrod, 1907 chng minh rng bnh ny do mt gene ln, thng xy ra cc gia nh c ngi cng dng mu ly nhau. Biu hin ca bnh ny rt d nhn bit: nc tiu ca bnh nhn c mu en do trong nc tiu cha mt lng ln axit homogenetizic. ngi bnh thng, axit homogenetizic s c enzyme oxydase phn gii thnh axit axetoaxetic. Khi thiu enzyme oxydase th axit homogenetizic khng c phn gii v thi ra theo nc tiu, b oxy ho trong khng kh to thnh mt hp cht c mu en. Khi cn b tt bm sinh ny cha gy hu qu nghim trng nhng khi ng tui b vim khp nng, sn tch lu nhiu sc t. 1.2. Cc bnh nhim sc th Nh s pht trin ca cc phng php nghin cu di truyn hc ngi, c bit l phng php di truyn t bo lm r c ch pht sinh nhiu loi bnh nhim sc th, do s phn ly khng bnh thng ca nhim sc th trong qu trnh phn bo hoc do d dng, sai hnh nhim sc th t cc ri lon trong cu trc, hnh thi. 1.2.1. Hi chng Down Hi chng ny c Langdom Down pht hin ln u tin vo nm 1866. Tn s bt gp khong 1/700 tr s sinh v l bnh nhim sc th bt gp cao nht. Thng th b nhim sc th ca bnh nhn c 47 chic, tha mt nhim sc th 21. Trong mt s trng hp hi chng ny cn l kt qu ca chuyn on gia nhim sc th 21 vi nhim sc th ca nhm D hoc nhm G. Biu hin bnh l l ngu n bm sinh, gim tr lc, nhiu d tt cc c quan ni tng, khng c kh nng sinh dc, vc dng b, ln, c rt, u b, chm dt, mt trn, khe mt xch, mi dy, li dy c xu th th ra thng xuyn...S liu v hi chng Down tr em Vit Nam c trnh by trong chng 3. 1.2.2. Hi chng Klinefelter H. F.Klinefelter m t hi chng ny ln u tin vo nm 1942. Tn s bt gp l khong 1/1000 b trai sinh ra. Kiu nhn ca bnh nhn l 47, XXY. Nguyn nhn c th l do th thai t mt t bo trng XX vi tinh trng Y hoc t mt trng X vi tinh trng XY. Bnh nhn l nam khng bnh thng v tuyn sinh dc, c mt s nt ging n , c v nh n, c bit l cc tnh trng gii tnh th cp, tr tu km pht trin, khng c con, kh ngi cao, chn tay di (Hnh 11.4).

1.2.3. Hi chng Turner Hi chng ny c H.H.Turner pht hin ln u tin vo nm 1938. Tn s bt gp khong 1/3000 tr em gi s sinh. Kiu nhn ca bnh nhn l 45, X. Nguyn nhn l do mt t bo trng hoc mt tinh trng khng c nhim sc th gii tnh, hoc do mt i mt nhim sc th gii tnh trong nguyn phn nhng ln phn ct u tin sau khi hnh thnh hp t XX hoc XY. Bnh nhn c tm vc b, thng b cc d hnh nh khng c bung trng, thiu cc tnh trng gii tnh th cp do khng c con v nhiu d dng b ngoi khc (Hnh 11.5).
Hnh 11.4 Hi chng Klinefelter
Hnh 11.5 Hi chng Turner

Cn ni thm l, Vit Nam, c cc s liu nghin cu c cng b v cc bt thng nhim sc th trong cc th bnh Leukemia cp, mt dng bnh tng bch cu, hoi huyt (Phm Quang Vinh, 2003). 2. C s di truyn ung th C mt s bng chng r rng cho thy rng ung th l mt bnh do s bin i ca gene gy ra. Cc t bo khi u c s lng nhim sc th bin i v cc nhim sc th thng c s sp xp li. Mt vi chuyn on l c trng vi cc dng ung th nht nh. Phn ln cc tc nhn t bin l cc tc nhn gy ung th. T bm (predisposition) vi cc dng ung th c tm thy l di truyn mt s gia nh. C ba loi gene lin quan n ung th c pht hin l gene ung th (oncogene), gene c ch khi u (tumor suppressor gene) v gene sa cha DNA (DNA repair gene). Phn ln gene ung th c ngun gc t protooncogene. Protooncogene l cc gene m ho cc sn phm kim sot s sinh trng v bit ho ca t bo. Protooncogene c hot ho to thnh gene ung th do t bin hoc biu hin qu mc. Ln u tin chng c pht hin trong cc retrovirus gy ung th, nhng cng c th c pht hin trong cc khi u khng c virus bng s chuyn nhim vo trong cc t bo 3T3 ca chut. Gene ung th cc retrovirus l v-onc v khi trong cc khi u l c-onc. Thnh thong gene ung th c khuych i trong cc t bo khi u. Phn ln cc gene ung th hot ng nh allele tri lm tng chc nng t bin dn n loi b kh nng iu ho ca s kim sot chu trnh t bo. Ngc li vi gene c ch khi u, gene ung th khng biu hin cc t bin dng mm (germline mutations) m cc t bin ny gy nn cc hi chng ung th di truyn (inherited cancer syndromes). Trong khi t bin soma gy ra ung th n pht (sporadic cancer). Mt s khi u do chuyn on c hiu lm tng s phin m ca gene ung th c m t c im. Hin tng ny c tm thy u lympho ca Burkitt (Burkitts lymphoma).Hoc s dung hp ca hai protooncogene c th to thnh mt gene mi l vi cc c tnh ung th c quan st bnh bch cu dng tu mn tnh (chronic myeloid leukemia). Gene c ch khi u cn n c hai bn sao tr nn bt hot trc khi cc khi u c th pht trin. Cc t bin ny l t bin ln. Thng thng s bt hot xy ra l do t bin im mt allele v mt mt allele khc. S bt hot ca gene c ch khi u xy ra cc t bo soma trong qu trnh pht trin ca khi u. Gene c ch khi u c nghin cu k nht l gene p53. Trong hn 40% cc khi u ca ngi, gene ny b t

bin. p53 ng vai tr iu ho chu trnh t bo v qu trnh cht theo chng trnh ca t bo (apoptosis) Mt s ung th him gp v mt phn nh ung th ph bin tp hp li trong mt s gia nh. Cc gia nh ny c gi l c t bm di truyn (hereditary predisposition) vi ung th. T bm di truyn ny c th tr thnh mt dng c trng ca ung th v d nh ung th v hoc mt loi ung th khc. T bm di truyn c th tng ln thng qua cc t bin dng mm ca gene c ch khi u. Tt c cc t bo soma ca mt c th mang mt allele t bin v c nguy c hnh thnh khi u tng cao do s bt hot ca mt allele n bnh thng. V d ngi gm c gene vng mc (retinoblastoma -Rb) nm trn nhim sc th 13 v gene t bm di truyn ung th v v ung th bung trng (BRCA1) nm trn nhim sc th 17. Allele bnh thng b bt hot trong khi u. Cc gene ung th gia nh thng b t bin soma trong cc trng hp khng thng xuyn (khng di truyn) ca cng loi ung th. Ung th di truyn cho thy kiu di truyn do gene tri trn nhim sc th thng bi v ngi bnh truyn t bm di truyn cho mt na s con ci ca h. Tuy nhin cc t bin gene c ch khi u l t bin ln. N c gii thch bng thc t l nu mt c th ch c mt bn sao hot ng chc nng ca gene c ch khi u trong mi t bo th n rt thch hp cho vic mt t bin th hai s gy bt hot bn sao hot ng chc nng bnh thng ny trong mi t bo. Sau cc t bo t bin kp ny c th pht trin thnh mt khi u. Cc gene sa cha DNA cng kt hp vi ung th di truyn. V d bnh da sng ho sc t (xeroderma pigmentosum) do mt t bin ln trn nhim sc th thng. Ngi bnh rt nhy cm vi nh sng mt tri bi v h thiu endonuclease xc tc cho vic sa cha DNA. Cc allele t bin ca mt s gene khc c th dn n t bm ung th rut kt (colon cancer). Hai trong s cc t bin ny cng lin quan n s sa cha cc DNA khng hp i (DNA mismatches). Cc t bo khi u sa cha DNA tn thng t hiu qu hn cc t bo bnh thng v to ra cc t bin vi tn s cao v mt s cc t bin s tr thnh cc gene c ch khi u hoc protooncogene. y l kiu hnh ca gene tng cng t bin (mutator phenotype).
VI. T vn di truyn y hc (Medical genetic counseling)

T vn di truyn y hc hay cn gi l li khuyn di truyn, l s trao i kin, gii thch v nguyn nhn, c ch v mt bnh tt di truyn no v cho li khuyn v kh nng mc bnh i con ca cc cp v

chng m bn thn h hoc mt s ngi trong dng h mc phi bnh y. Li khuyn di truyn nhm mc ch gip cc cp v chng n xin li khuyn di truyn t quyt nh c nn sinh con hay khng v nu sinh th phi nh th no trnh cho ra i cc a tr tt nguyn gim gnh nng cho gia nh v x hi, cng nh cung cp cho h cc phng php, bin php phng, iu tr v hn ch cc hu qu cho bn thn v cho con ci. c c li khuyn di truyn chnh xc, nh t vn di truyn y hc phi tm hiu tin s gia nh, theo di ph h, xc nh ng l bnh di truyn v y l t bin tri hay ln v di truyn theo qui lut no, thm khm cho ngi b bnh v trong mt s trng hp phi thm khm cho c nhng ngi c lin quan trong gia nh, lm cc xt nghim chn on. Cc i tng cn xin li khuyn di truyn l cc cp v chng c con ci b bnh di truyn; cc nam n thanh nin trc khi xy dng gia nh khi m mt trong hai gia nh hoc c hai gia nh c ngi mc bnh di truyn; nhng ngi b v sinh, sy thai lin tc,...; cc cp v chng mun sinh con theo mun; c hai v chng hoc ch v hay chng lm vic trong iu kin c hi; bit v hoc chng mang gene bnh; cc cp v chng ln tui, c bit l ngi v v nhng ngi kt duyn cng dng h... Trc y li khuyn di truyn hon ton da vo s hiu bit v qui lut di truyn, tnh xc sut xy ra cho tng trng hp c th, phn tch ph h xc nh bnh di truyn theo qui lut no...Ngy nay k thut chn on trc sinh c thc hin vi nhng phng php phn tch ng tin cy ngay khi c th cn giai on phi thai trong c th m nh phng php chc d dch i phn tch di truyn hc t bo v ho sinh, quan st gin tip bo thai nh siu m, quan st trc tip bo thai bng soi phi thai, ly cc mu sinh phm t phi thai (mu, tua nhau thai...) xt nghim, phng php nh lng AFP (alpha feto protein) trong mu m. Chn on trc sinh cho php nh gi tnh trng ca b nhim sc th, ca b gene, tnh trng hot ng ca cc enzyme trong bo thai gip t vn di truyn y hc c c s khoa hc chc chn cho li khuyn chnh xc.
Cu hi v Bi tp
1. Mt cp v chng c mt a con b bnh u x nang. y l mt bnh him gp, c quy nh bi mt t bin lndi truyn theo nh lut Mendel. Cp v chng ny d nh sinh thm con. Hy tnh xc sut a cob tip theo: (a) Mang gene bnh; (b) Khng b bnh.

2. ngi, tnh trng tc qun tri so vi tnh trng tc thng, do gene nm trn nhim sc th thng quy nh; cn bnh m mu do gene ln nm trn nhim sc th gii tnh X gy nn. B v m c tc qun, mt bnh thng sinh ra mt con trai tc qun, m mu. Hy xc nh kiu gene ca b m. 3. B m u c nhm mu A, sinh con trai c nhm mu A, con gi c nhm mu O. Hy tm kiu gene chc c ca nhng ngi trong gia nh trn. 4. Kt qu thng k trn mt s lng ln cc a tr sinh ra t nhiu cp v chng, trong cc ngi chng u b bnh xn men rng cn cc ngi v c men rng bnh thng cho thy: 50% b bnh xn men rng u l gi : 50% c men rng bnh thng ton l trai. Hy xc nh tnh cht di truyn ca bnh xn men rng v vit s lai t P n F1. Cho bit tnh trng men rng do mt gene quy nh. 5. Chng bnh thng, ly v b bnh mu kh ng. Con ca h c b bnh mu kh ng khng? 6. "Bnh m mu v bnh mu kh ng l bnh ca nam gii". Quan nim nh vy c ng khng? Hy gii thch ti sao. 7. Trnh by cc phng php nghin cu di truyn ngi. 8. Trnh by cc phng php lp bn di truyn ngi. 9. Nu c im di truyn ca cc gene tri-ln trn nhim sc th thng v nhim sc th gii tnh. 10. Trnh by mt s bnh di truyn do ri lon chuyn ho v cc bnh nhim sc th.
Ti liu Tham kho

Ting Vit Trn Quc Dung. 2003. Bi ging Di truyn hc ngi. Trng i hc S phm Hu, Hu. Phan C Nhn, Nguyn Minh Cng, ng Hu Lanh. 1999. Di truyn hc ngi. Trong: Di truyn hc (Phan C Nhn , ch bin). Tp II. Trang: 161-248. NXB Gio Dc. Ting Anh Doris Bachtrog, Brian Chalesworth. 2001. Towards a complete sequence of the human Y chromosome. Genome Biol 2 (5): reviews 1016.1- reviews 1016.5. Published online 2001 April 26.

Jorde, Carey, Bamshad, White. 2000. Medical Genetics. Printed in the United States of America. Winter P. C., Hickey G.I., Fletcher H. L. 1998. Genetics. Printed by Biddles Ltd, Guilford, UK. http://users.rcn.com/jkiball.ma.ultranet/BiologyPages/S/SexChromosomes. html http://cas.bellarmine.edu/tietjen/HumanBiology/bill_developmental_abnor malities.htm

Chng 12
Di truyn hc Qun th
Ln u tin vo nm 1908, G.Hardy v W.Weinberg chng minh rng tnh di truyn t n khng lm thay i tn s allele trong qun th, sau ny c gi l nguyn l Hardy-Weinberg (Hardy-Weinberg principle). N t nn mng cho di truyn hc qun th (population genetics) - mt nhnh ca di truyn hc - nghin cu thnh phn di truyn ca cc qun th sinh vt v cc qu trnh nh hng ln cc tn s gene ca chng. Tuy nhin, n u thp nin 1930 lnh vc nghin cu ny mi thc s pht trin nh cc cng trnh v i ca R.A.Fisher, J.B.S. Haldane v S.Wright m thc cht l cc m hnh ton hc. T di truyn hc qun th tr thnh nn tng ca cc thuyt tin ho hin i. Trong chng ny, chng ta s tho lun mt s khi nim c bn ca di truyn hc qun th, nguyn l Hardy-Weinberg v mi quan h gia cc tn s allele v kiu gene trong cc trng hp khc nhau, ni phi v s gia tng tn s ca cc th ng hp, v tm hiu s lc vai tr ca cc nhn t tc ng ln thnh phn di truyn qun th.
I. Cc khi nim c bn ca Di truyn hc qun th

1. Qun th (population) Trong tin ho, c th khng c xem l n v thch hp bi v: kiu gene ca mt c th c gi nguyn trong qung i ca n; hn na, c th c tnh tm b (d n c th sng ti c nghn nm nh cy tng...). Ngc li, mt qun th th c tnh lin tc qua thi gian v mt khc, thnh phn di truyn ca n c th thay i tin ho qua cc th h. S hnh thnh cc qun th a phng ti nhng vng lnh th khc nhau chnh l phng thc thch ng ca loi trc t nhin. Qun th v vy c xem l n v tin ha c s. Theo A.V.Yablokov (1986), qun th l mt nhm cc c th cng loi c kh nng giao phi t do vi nhau, chim c mt khu phn b xc nh v tri qua mt khong thi gian tin ho lu di hnh thnh nn mt h thng di truyn c lp v mt sinh thi ring. Ni ngn gn, qun th l mt nhm sinh vt c kh nng giao phi qua li v cng chia x mt vn gene chung (Ridley 1993). N cn c gi l qun th Mendel, m tp hp ln nht l loi (species). 2. Cc h thng giao phi (mating systems) Trn nguyn tc, cu trc di truyn ca qun th th h sau c xc nh bi xc sut kt hp ca cc giao t th h trc trong qu trnh th

tinh. Do , n ph thuc vo kiu giao phi ca cc b m. Trong di truyn hc qun th, ngi ta phn bit ba kiu giao phi: Giao phi ngu nhin hay ngu phi (random mating hay panmixia), giao phi chn la (assortative mating), v ni phi (inbreeding). - Ngu phi l kiu giao phi trong xy ra s bt cp ngu nhin gia cc c th c v ci trong qun th. Lu rng nh ngha qun th trn y c p dng cho cc qun th thuc h thng ngu phi; chng chim v tr rt quan trng trong h thng cc loi v c cp ch yu trong sut ch ny. - Giao phi chn la l kiu giao phi trong cc c th c v ci khng bt cp ngu nhin m c s la chn theo kiu hnh. C hai trng hp: (1) Nu nh cc c th c xu hng giao phi vi cc c th khc c kiu hnh tng t, th gi l giao phi chn la dng tnh (positive assortative mating); v (2) Nu nh s la chn t c quan tm nhng tn s ca cc cp giao phi vn khc xa vi tn s ca cc cp ngu phi, th gi l giao phi khng la chn (disassortative mating) hay chn la m tnh (negative assortative mating). Chng hn, ngi, s giao phi c la chn xy ra i vi cc tnh trng nh chiu cao, mu mt, mu tc...V vy n ch nh hng n cc tn s kiu gene ca locus no c lin quan n vic xc nh kiu hnh c s dng trong giao phi. Cn kiu giao phi khng la chn ph bin trong cc h thng t bt dc (self-sterility) thc vt. - Ni phi l s giao phi khng ngu nhin xy ra gia cc c th c quan h h hng gn hoc in hnh l s t th tinh (xem mc IV). 3. Vn gene (gene pool) Vn gene l tp hp ton b cc allele tt c cc gene ca mi c th trong qun th ti mt thi im xc nh. Vn gene ny c s dng chung cho cc c th trong qun th. Mi qun th c trng bng mt vn gene nht nh v n c m t bng tn s cc allele tng locus. 4. Tn s kiu gene v tn s allele m t thnh phn di truyn ca mt qun th ta cn phi xc nh kiu gene ca cc c th v s c th ca mi kiu gene. Gi s trong mt qun th sinh vt lng bi gm N c th, xt mt locus A thuc nhim sc th thng (autosome) vi hai allele A1 v A2 c mt trong cc c th. Lc s c ba kiu gene: A1A1, A1A2 v A2A2 vi s lng tng ng l N11, N12 v N22; (N = N11 + N12 + N22). Nu k hiu P, H v Q l tn s tng ng vi cc kiu gene trn, ta c:

P = N11 / N; H = N12 / N v Q = N22 / N ; (P + H + Q = 1) T y ta c th tnh c cc tn s gene hay allele (gene or allelic frequencies) A1 v A2, vi k hiu tng ng l p v q ( p +q =1), nh sau: 2 N 11 + N 12 1 p= =P+ H 2N 2 1 2 N 22 + N 12 q= =Q+ H ( hay q =1-p ) 2 2N Tm tt: (1) Vn gene ca mt qun th c N c th bao gm 2N h gene n bi. Mi h gene gm tt c cc thng tin di truyn nhn c t mt cha m. i vi mi locus autosome, trong vn gene qun th s c 2N allele. (2) Tn s kiu hnh (phenotypic frequency) bng s lng c th ca kiu hnh c th chia cho tng s c th ca qun th. (3) Tn s kiu gene (geneotypic frequency) bng s lng c th ca kiu gene c th chia cho tng s c th ca qun th. (4) Tn s allele (allelic frequency) bng hai ln s lng c th ng hp cng vi s c th d hp v allele chia cho hai ln tng s c th ca qun th; hay tn s ca mt allele bng tn s kiu gene ng hp cng vi mt na tn s kiu gene d hp v allele . Lu : (i) Tng cc tn s kiu hnh, kiu gene hay allele thuc mt locus no lun lun bng n v; (ii) Tn s allele hay tn s gene nh mt s nh khoa hc thng gi (Crow 1986; Falconer v Mackay 1996) l khi nim cn bn nht ca di truyn hc qun th; n l du hiu c trng ca mt qun th cho php phn bit vi cc qun th khc trong cng mt loi; (iii) cho tn s cc allele quan st c l c trng ca mt qun th, th mu thu c phi l ngu nhin v c kch thc ln; (iv) tin cho mt s mc ch m t cc bin d di truyn mt locus, ngi ta s dng ch yu tn s cc allele ch khng phi tn s cc kiu gene, bi v mt locus thng c s allele t hn s kiu gene; (v) Thut ng "thnh phn" hay "cu trc di truyn ca qun th" dng ch tn s tng i ca cc allele v cc kiu gene trong qun th ti mt thi im xc nh. V d: S liu phn b ca h nhm mu M-N mt s qun th ngi bng 12.1 cho thy: (1) Mi qun th mt vng a l nht nh u c cc tn s allele c trng ; (2) Trong khi qun th ngi M gc u c cc tn s allele M v N hu nh tng ng, mc d tn s allele M cao hn khong 8%, th qun th th dn c c tn s allele N rt cao

(nhiu gp 4,6 ln tn s allele M); cn b tc da Navaho ni ring v vng Trung-Nam M ni chung c tn s allele M rt cao.
Bng 12.1 Tn s cc nhm mu h M-N mt s qun th ngi
Qun th Btc Navaho S lng Tn s kiu gene Tn s allele
M 305 22
MN 52 216
N 4 492
L L
M M
L L
M N
L L
N N
LM
LN
0,845 0,030 0,292
0,144 0,296 0,496
0,011 0,674 0,213
0,917 0,083 0,178 0,822 0,539 0,461
Th dn c M gc u
1787 3039 1303
II. Nguyn l Hardy-Weinberg v trng thi cn bng ca qun th

1. Nguyn l Hardy-Weinbeirg Nm 1908, nh ton hc ngi Anh Godfrey H.Hardy v bc s ngi c Wilhelm Weinberg c lp chng minh rng c tn ti mt mi quan h n gin gia cc tn s allele v cc tn s kiu gene m ngy nay ta gi l nh lut hay nguyn l Hardy-Weinberg (vit tt: H -W ). 1.1. Ni dung nguyn l H-W Trong mt qun th ngu phi kch thc ln, nu nh khng c p lc ca cc qu trnh t bin, di nhp c, bin ng di truyn v chn lc, th tn s cc allele c duy tr n nh t th h ny sang th h khc v tn s cc kiu gene (ca mt gene gm hai allele khc nhau) l mt hm nh thc ca cc tn s allele, c biu din bng cng thc sau: ( p + q )2 = p2 + 2pq + q2 = 1 1.2. Chng minh mt qun th Mendel, xt mt locus autosome gm hai allele A1 v A2 c tn s nh nhau c hai gii c v ci. K hiu p v q cho cc tn s allele ni trn (p + q =1). Cng gi thit rng cc c th c v ci bt cp ngu nhin, ngha l cc giao t c v ci gp g nhau mt cch ngu nhin trong s hnh thnh cc hp t. Khi tn s ca mt kiu gene no chnh l bng tch ca cc tn s hai allele tng ng. Xc sut mt c th c kiu gene A1A1 l bng xc sut (p) ca allele A1 nhn t m nhn vi xc sut (p) ca allele A1 nhn t b, hay p.p = p2. Tng t, xc sut m mt c th c kiu gene A2A2 l q2. Kiu gene A1A2 c th xut hin theo hai cch: A1 t m v A2 t b vi tn s l pq, hoc A2 t m v A1 t b cng vi tn s pq; v vy tn s ca A1A2 l pq + pq = 2pq (Bng 12.2). iu chng minh trn c tm tt nh sau:

* Qun th ban u c 3 kiu gene : A1A1 A1A2 A2A2 Tng Tn s cc kiu gene : P H Q Tn s cc allele : p = P + H ; q = Q + H 1 * Qun th th h th nht sau ngu phi c : Tn s cc kiu gene = (p + q)2 = p2 + 2pq + q2 1 2 Tn s cc allele: f(A1) = p + (2pq) = p(p+q) = p f(A2) = q2 + (2pq) = q(p+q) = q Nhn xt: T chng minh trn cho thy cc tn s allele th h con ging ht th h ban u, ngha l f(A1) = p v f(A2) = q. Do , cc tn s kiu gene th h tip theo vn l p2, 2pq v q2 (ging nh th h th nht sau ngu phi). iu chng t rng cc tn s kiu gene t c cn bng ch sau mt th h ngu phi. Trng thi n nh v thnh phn di truyn c phn nh bng cng thc H-W nh vy c gi l cn bng H-W (Hardy-Weinberg equilibrium).
Bng 12.2 Cc tn s H-W sinh ra t s kt hp ngu nhin cc giao t
Tn s giao t ci
p(A1)
Tns gt c
q(A2) pq(A1A2) q2(A2A2)
p(A1) q(A2)
p2(A1A1) pq(A1A2)
1.3. Cc mnh v h qu (1) Nu nh khng c p lc ca cc qu trnh tin ho (t bin, di nhp c, bin ng di truyn v chn lc), th cc tn s allele c gi nguyn khng i t th h ny sang th h khc. y l mnh chnh ca nguyn l hay nh lut H-W. (2) Nu s giao phi l ngu nhin, th cc tn s kiu gene c quan h vi cc tn s allele bng cng thc n gin: ( p+q )2 = p2 + 2pq + q2 =1. (3) H qu 1: Bt lun cc tn s kiu gene ban u (P, H, Q) nh th no, min sao cc tn s allele hai gii l nh nhau, ch sau mt th h ngu phi cc tn s kiu gene t ti trng thi cn bng (p2, 2pq v q2). (4) H qu 2: Khi qun th trng thi cn bng th tch ca cc tn s ng hp t bng bnh phng ca mt na tn s d hp t, ngha l: (2 pq ) 2 p2.q2 = [ ] 2

Tht vy, khi qun th trng thi cn bng l tng, ta c: H = 2pq Bin i ng thc trn ta c: pq = H Bnh phng c hai v, ta c: p2.q2 = (H)2, trong H = 2pq. Nh vy ng thc ny cho thy mi tng quan gia cc thnh phn ng hp v d hp khi qun th trng thi cn bng l tng. (5) H qu 3: (i) Tn s ca cc th d hp khng vt qu 50%, v gi tr cc i ny ch xy ra khi p = q = 0,5 H = 2pq = 0,5; lc ny cc th d hp chim mt na s c th trong qun th; (ii) i vi allele him (tc c tn s thp), n chim u th trong cc th d hp ngha l, tn s th d hp cao hn nhiu so vi tn s th ng hp v allele . iu ny gy hu qu quan trng i vi hiu qu chn lc (xem thm mc 1.5.2 di y). 1.4. Tn s giao phi v s kim chng nguyn l H-W Nguyn l H-W c th c chng minh theo mt cch khc da trn tn s ca cc kiu giao phi. Mc d n cng knh hn phng php xt nhng li cho thy r hn bng cch no cc tn s H-W pht xut t quy lut phn ly ca Mendel. Xt cu trc giao phi ca qun th ngu phi nh trn ta thy c c thy l chn kiu giao phi vi tn s giao phi nh Bng 12.3. V tn s mi kiu gene hai gii c xem l nh nhau, nn mt s kiu giao phi thun nghch l tng ng v vy ch cn li su kiu giao phi khc nhau vi tn s tng ng c nu hai ct u tin ca bng 12.4. By gi ta xt cc kiu gene i con sinh ra t mi kiu giao phi v sau tm tn s ca mi kiu gene trong ton b i con, vi gi thit rng tt c cc kiu giao phi u hu th ngang nhau v tt c cc kiu gene u c sc sng nh nhau. Kt qu ny c trnh by pha bn phi Bng 12.4. Sau khi rt gn ta c cc tn s kiu gene i con tng ng l p2 , 2pq v q2 ( dng cui cng ca bng). Cc tr s ny chnh l cc tn s cn bng H-W (equilibrium frequencies) t c sau mt th h ngu phi, bt lun cc tn s kiu gene i b m nh th no.
Bng 12.3 Tn s ca cc kiu giao phi ngu nhin
Gii c Gii ci
A1A1 (P) P2 PH PQ
A1A2 (H) PH H2 QH
A2A2 (Q) PQ QH Q2
A1A1 (P) A1A2 (H) A2A2 (Q)

Bng 12.4 Nguyn l Hardy-Weinberg i vi hai allele
B m Kiu giao phi Tn s i con
A1A1 P2 PH H2
A1A2 PH 2PQ H2 HQ
A2A2 H2 HQ Q2
A1A1 A1A1 A1A1 A1A2 A1A1 A2A2 A1A2 A1A2 A1A2 A2A2 A2A2 A2A2 Tng
P2 2PH 2PQ H2 2HQ Q2 1
(P+H)2 =p2 : 2(P+H)(Q+H) =2pq : (Q+H)2 = q2
2. Nhng ng dng ca nguyn l Hardy-Weinberg 2.1. Xc nh tn s ca allele ln Trong trng hp tri hon ton, ta khng th phn bit cc th d hp vi th ng hp tri. V vy, trn nguyn tc, ta khng th tnh c cc tn s allele. Tuy nhin, c th gi nh cc tn s kiu gene dng cn bng, qua tnh c tn s allele ln v d on tn s ca cc kiu gene trong qun th. Chng hn, bch tng (albinism) ngi l tnh trng ln tng i him gp. Nu nh k hiu A cho allele xc nh sc t bnh thng v a cho allele bch tng, kiu gene ca ngi b bch tng l aa, trong khi nhng ngi bnh thng th hoc l AA hoc l Aa. Gi s trong mt qun th ngi tn s ca nhng ngi b bch tng l 1/10.000. Theo nguyn l H-W, tn s ca th ng hp ln l q2 = 0,0001 nn q = f (aa) = 0,0001 = 0,01. Do tn s ca allele A l: p = 1 0,01 = 0,99 (v p + q = 1). T y xc nh c tn s ca hai kiu gene cn li: f(AA) = p2 = (0,99)2 = 0,9801 (hay ~98%) f(Aa) = 2pq = 2(0,99)(0,01) = 0,0198 (hay ~ 2%) Lu trong trng hp tn s allele ln l rt thp, ngha l kch thc mu ln, ta cn phi ly s thp phn y m bo chnh xc cho cc kt qu tnh ton sau cng. 2.2. Xc nh tn s ca cc "th mang" (carrier) Mt iu l th ca nguyn l H-W l ch, cc allele him ni chung l cc allele ln gy bnh trong qun th thng n tng trong cc th d hp (gi l th mang) v ta c th tnh c tn s ca chng nu nh bit c tn s allele. Nu cho rng c s cn bng H-W th tn s ca cc th mang allele bnh ln trong qun th c c tnh l H = 2q(1-q).

V tn s ca cc th d hp trong s nhng c th bnh thng, k hiu H, l t s f(Aa)/ f(AA+Aa), trong a l allele ln vi tn s q. Khi : 2q 2 pq 2q (1 q) = = H = 2 2 p + 2 pq (1 q) + 2q(1 q ) 1 + q V d: Vi trng hp bch tng ni trn, tn s ca aa l 0,0001 th tn s ca nhng ngi d hp (Aa) l 0,02 , ngha l trong 50 ngi c mt ngi mang allele bch tng. y l mt t l rt cao! Mt khc, tn s allele a nhng ngi d hp l 0,02: 2 = 0,01 trong khi nhng ngi bch tng l 0,0001, nh vy allele a nhng ngi d hp c nhiu hn nhng ngi bch tng khong 100 ln (0,01 : 0,0001 = 100 ). Tng qut, nu tn s ca mt allele ln trong qun th l q, th s c pq allele ln trong cc th d hp v q2 allele ln trong cc th ng hp. T s y l pq/q2 = p/q, v nu nh q rt b th t s s xp x 1/q. Nh vy, khi tn s ca mt allele ln cng thp bao nhiu, th t l ca allele trong cc th d hp cng cao by nhiu. Tng t, c th ly nhiu v d v cc allele ln gy bnh ngi. in hnh l bnh ri lon chuyn ho c tn l phenylxetn-niu (phenylketonuria = PKU) do mt allele ln n, c th pht hin sm vi ngy sau sinh. Mt kt qu iu tra Birmingham trong hn ba nm cho thy c 5 trng hp b bnh trong s 55.715 b (Raine v cs 1972). Tn s cc th ng hp ln xp x 1/11.000 hay 90 x 10-6. Tn s allele ln l q = 90 x10 6 = 0,0095. Tn s cc th d hp trong c qun th (H = 2pq) v trong s cc th bnh thng (H= 2q/1+q) u xp x bng 0,019. Nh vy khong 2% s ngi bnh thng l c mang mm bnh PKU. Cc kt qu ny tht ng ngc nhin: bng cch no cc th d hp v allele ln li ph bin n nh vy, trong khi tn s bnh thc t l qu thp! n y ta c th khng nh rng: Nu nh ai c tng mun loi b mt allele ln him gy bnh no ra khi qun th hng ci thin chng tc chng hn, qu l khng tng! Tht vy, nu gi t l s th h cn thit bin i tn s allele ban u l q0 xung cn qt th h th t, ta c t =1/qt -1/q0. Gi s q0 = 0,01, mun gim xung cn 0,001 phi cn ti 900 th h; tng t, gim tn s xung cn 0,0001 phi cn n 9.900 th h. Th tng tng ngi mt th h trung bnh l 30 nm, thi gian y ln n dng no (9.900 x 30 = 297.000 nm)! 2.3. Kho st trng thi cn bng ca qun th T nguyn l H-W v cc h qu rt ra c trn cho php ta vn dng xc nh xem cu trc di truyn ca mt qun th c trng thi

cn bng H-W hay khng. Di y ch lc trnh vi phng php tng qut i vi mt qun th ngu phi (Hong Trng Phn 2001), vi cc gi thit v k hiu c cp. Trc tin, cn nm vng nguyn tc ny trong suy lun: Theo nguyn l H-W, cc tn s kiu gene i con c xc nh nh tn s allele b m chng. Nu qun th trng thi cn bng, tn s cc allele s nh nhau c hai th h, v vy tn s allele quan st c i con c th dng y nh th n l tn s allele i b m tnh cc tn s kiu gene k vng theo nguyn l H-W. Nh vy, v nguyn tc, mt qun th c coi l trng thi cn bng nu nh n tha mn mt trong nhng kh nng sau y; ngc li, qun th khng trng thi cn bng. (1) Cc tn s kiu gene quan st c (P, H v Q) phi xp x bng cc tn s k vng tng ng (p2, 2pq v q2), ngha l thnh phn di truyn ca qun th phi tho mn cng thc H-W. V mt s lng, qun th c coi l trng thi cn bng nu nh c s ph hp st sao gia cc con s quan st v k vng i vi mi kiu gene, ngha l: N11 p2N ; N12 2pqN; v N22 q2N. (2) Tn s th d hp quan st phi xp x bng tn s k vng (H 2pq), ngha l: p.q H hay P.Q (H)2 (3) Tn s ca mi kiu gene quan st c gia hai th h lin tip l tng ng nhau. Nu ta gi tn s ca cc kiu gene A1A1, A1A2 v A2A2 tng ng th h th nht l P1, H1v Q1 v th h th hai l P2, H2 v Q2, lc : P1 P2 ; H1 H2; v Q1 Q2. (4) i vi trng hp kho st cn bng H-W hoc giao phi ngu nhin da trn tn s giao phi hoc s lng cp giao phi ca cc kiu giao phi khc nhau, ta c th so snh nh sau:
Kiu giao phi Tn s Quan st K vng S lng Quan st
K vng
p2.p2.N/2 2(p2)(2pq)N/2 2(p2)(q2)N/2 (2pq)(2pq)N/2 2(2pq)(q2)N/2 q2.q2.N/2 N/2
A1A1 A1A1 A1A1 A1A2 A1A2 A2A2 Tng
x x x x x x
A1A1 A1A2 A2A2 A1A2 A2A2 A2A2
P 2PH 2PQ H2 2QH Q2 1
p .p 2(p2)(2pq) 2(p2)(q2) (2pq)(2pq) 2(2pq)(q2) q2.q2 1
P .N/2 2P.H.N/2 2P.Q.N/2 H2.N/2 2Q.H.N/2 Q2.N/2 N/2

(5) Phng php Khi-bnh phng (Chi-square method) Khi so snh gia cc s liu quan st v k vng thng c th c s sai lch khng ng k hoc ng k. V ranh gii phn nh gia chng l khng r rng khin ta kh m khng nh qun th trng thi cn bng hoc khng. Trong trng hp , ta phi s dng phng php 2 (xem chng 1). V d: kho st trng thi cn bng H-W, ta hy xt qun th ngi M da trng gc u cho bng 12.1. T s ngi mang cc nhm mu M, MN vN tng ng l 1.787; 3.039; v 1.303 (vi N = 6.129), ta tnh c tn s ca cc allele M v N l p v q nh sau: p = 1.787 + 1/2(3.039) = 0,539 v q = 1 - p = 0,461. T y tnh c tn s k vng ca cc kiu gene: MM p2 = (0,539)2 = 0,292 MN 2pq = 2(0,539)(0,461) = 0,497 NN q2 = (0,461)2 = 0,211 V s c th k vng ca chng: MM p2 N = 0,292 6.129 = 1.787,2 MN 2pq N = 0,497 6.129 = 3.044,9 NN q2 N = 0,211 6.129 = 1.296,9 So snh cc s liu quan st v k vng v tng kiu gene ta thy c s ph hp st sao, chng t qun th trng thi cn bng H-W. Tht vy, nu kim tra bng trc nghim 2, ta c: 2 =
(1787 1787,2) 2 (3039 3044,9) 2 (1303 1296,9) 2 + + = 0,04 1787,2 3044,9 1296,9
Tra bng phn phi 2 ng vi P = 0,05 v 1 bc t do ta tm c tr s 2 bng 3,84. V tr s thc t l rt nh so vi tr s l thuyt, chng t gia cc s liu quan st v k vng hu nh trng khp nhau hon ton; ngha l, qun th trng thi cn bng H-W.
III. M rng nguyn l Hardy-Weinberg

Sau y ta ln lt xt xem kh nng p dng nguyn l H-W vo ba trng hp: mt gene c nhiu hn hai allele (a allele), mt gene c hai allele vi tn s khc nhau hai gii tnh, v gene lin kt vi gii tnh. 1. a allele (multiple alleles) Vi qun th ngu phi nh ni trc, y ta ch thay gi thit

mt locus A c ba allele: A1, A2 v A3 vi tn s tng ng l p1, p2 v p3 (p1 + p2 + p3 = 1). Khi trong qun th c tt c su kiu gene vi s lng c th tng ng nh sau : Kiu gene : A1A1 A2A2 A3A3 A1A2 A1A3 A2A3 Tng S lng : N11 N22 N33 N12 N13 N23 N Theo nguyn tc, ta tnh c cc tn s allele: p1 = N11 + (N12 + N13) p2 = N22 + (N12 + N23) p3 = N33 + (N13 + N33) Bng cch lp bng t hp ngu nhin ca cc giao t v tn s ca chng, hoc bng cch khai trin bnh phng ca mt tam thc ta tnh c cc tn s cn bng H-W ch sau mt th h ngu phi nh sau: (p1 + p2 + p3)2 = p12 + p22 + p32 + 2p1p2 + 2p1p3 + 2p2p3 = 1 Tng qut, mt locus c n allele s c tt c n(n + 1)/ 2 kiu gene, trong gm n kiu ng hp v n(n 1)/2 kiu d hp. Tn ca mt allele bt k (pi) c tnh theo cng thc: pi = pii +
i j =1
Pij
trong pii - tn s kiu gene ng hp v pij- tn s kiu gene d hp. V d: Thng thng h nhm mu ABO c ly v d cho ba allele. V cc allele IA vIB l ng tri v allele IO l ln, nn trong qun th ngi bt k no cng s c bn nhm mu A, B, AB v O ng vi su kiu gene. tnh cc tn s allele trong trng hp ny ta phi gi nh qun th trng thi cn bng. t tn s ca cc allele IA, IB v IO ln lt l p, q v r (p + q + r =1). Khi ta tnh c tn s H-W ca cc nhm mu chnh l cc tn s quan st c (bng 12.5). Phng php tnh cc tn s allele nh sau: Trc tin, tn s allele IO (r) bng cc cn bc hai ca tn s nhm mu O (r2). Tn s ca hai allele cn li, p v q, c tnh bng cch kt hp tn s H-W ca mt nhm mu A hoc B vi nhm mu O theo mt trong hai phng php sau:
Phng php 1 Ta c f(A+0) = p +2pr + r = (p + r) p+r = p=
2 2 2
Phng php 2 V p +q +r = 1 q +r = 1 p Bnh phng 2 v ta c: (1 p)2 = (q + r)2 = f (B + O) 1p=
f ( A + 0) f ( A + O) r
Tng t, ta c :
f ( B + O)

q=
f ( B + O) r
p=1
f ( B + O) f ( A + O)
Tng t, ta c: q = 1
Mt cch tng i, ta c th tnh p hoc q ri suy ra ci cn li da vo tng p + q + r =1. Tuy nhin, nu tnh cn thn c ba tn s theo mt trong hai phng php trn ta s bit c tr s thc ca chng. Khi tng cc tn s allele tnh dc s khng ng bng n v mt cch chnh xc. iu ny c l gii l do t l cc kiu gene trong mu khng phi l cc t l H-W chnh xc v hn na, nhm mu AB khng c s dng trong tnh ton. V vy, khi kiu hnh khng c s dng n ( y l nhm mu AB) m c tn s cao hn th s mt mt thng tin s nghim trng hn, v phi cn n mt phng php chnh xc hn.
Bng 12.5 Tng quan gia cc nhm mu, kiu gene v tn s ca chng
Nhm mu Kiu gene Tn s K vng Quan st
A B O AB Tng
IAIA + IAIO IBIB + IBIO IOIO IAIB
p2 + 2pr q2 + 2qr r2 2pq 1
0,41716 0,08560 0,46684 0,03040 1,0
By gi ta hy xt mt mu nghin cu trn 190.177 phi cng vng quc Anh (UK) gm 79.334 A, 16.279 B, 88.782 O, v 5.782 AB ( Race v Sanger, 1954; dn theo Falconer 1989). Tng quan gia cc nhm mu, kiu gene v cc tn s ca chng c trnh by bng 12.5. p dng hai phng php trn ta tnh c cc tn s allele nh sau:
Tn s Phng php 1 Phng php 2
Allele
IA IB IO Tng
0,2569 0,0600 0,6833 1,0002
0,2567 0,0598 0,6833 0,9998

2. Tn s allele sai bit gia hai gii tnh Trn thc t, cc tn s allele nhim sc th thng hai gii tnh c th khc nhau. Chng hn, trong chn nui gia sc - gia cm tu theo mc tiu kinh t l ly sa, tht hoc trngm tng quan s lng c th c-ci s khc nhau. Khi vic p dng nguyn l H-W s nh th no? xt qun th ny, ta s dng k hiu v gi thit sau :
Allele Tn s Gii c Gii ci
A1 A2 Tng
p q 1
p q 1
Bng cch lp bng t hp ca cc giao t, ta xc nh c cu trc di truyn ca qun th sau mt th h ngu phi: (pA1 : qA2)(pA1 : qA2) = ppA1A1 : (pq+ pq) A1A2 : qqA2A2 R rng l n khng tha mn cng thc H-W. By gi n lt tn s cc allele ca qun th ny l nh sau: f(A1) = pp+ (pq+ pq) Thay gi tr q= 1 p, ta c: f(A1) = (p + p) Tng t: f(A2) = (q +q) t f(A1) = p v f(A2) = q , khi cu trc di truyn qun th th h tip theo s tho mn cng thc H-W: p2 A1A1 : 2pqA1A2 : q2A2A2. iu chng t rng, nu nh cc tn s allele (autosome) khi u l khc nhau hai gii, th chng s c san bng ch sau mt th h ngu phi v qun th t trng thi cn bng sau hai th h. V d: Mt qun th khi u c tn s cc allele A v a hai gii nh sau: p = 0,8; q= 0,2; p = 0,4; v q = 0,6. Nu nh ngu phi xy ra, th th h th nht c tn s cc kiu gene l: 0,32AA : 0,56Aa : 0,12aa. V tn s cn bng ca mi allele lc nh sau: p = (0,8 + 0.4) = 0,32 + (0,56) = 0,6 q = (0,2 + 0,6) = 0,12 + (0,56) = 0,4 th h th hai, qun th t cn bng vi cc tn s H-W l: 0,36AA : 0,48Aa : 0,16aa

3. Cc gene lin kt trn X Trong trng hp cc gene lin kt vi gii tnh, tnh hnh tr nn phc tp hn rt nhiu. gii ng giao t, mi quan h gia tn s allele v tn s kiu gene tng t nh mt gene autosome, nhng gii d giao t ch c hai kiu gene v mi c th ch mang mt allele. cho tin, ta coi gii d giao t l gii c. By gi ta xt hai allele A1 v A2 vi tn s tng ng l p v q, v t cc tn s kiu gene nh sau: Kiu gene: A1A1 A1A2 A2A2 A1 A2 Tn s : P H Q R S Theo nguyn tc, ta xc nh c tn s ca mt allele (v d A1): - gii ci (pc): pc = P + H - gii c (p): p = R - chung c qun th ( p ): p = pc + p Lu : Mi con ci c hai nhim sc th X v mi con c ch c mt X; v t l c : ci trn nguyn tc l 1:1, cho nn 2/3 cc gene lin kt gii tnh trong qun th l thuc v gii ci v 1/3 thuc v gii c. V vy, tn s ca cc allele A1 trong c qun th l: p = pc + p. R rng l cc tn s allele hai phn c v ci l khc nhau, do qun th khng trng thi cn bng. Trong khi tn s allele trong c qun th khng thay i qua cc th h, nhng s phn phi cc allele gia hai gii c s dao ng khi qun th tin dn n s cn bng. iu ny c chng minh nh sau. Theo quy lut lin kt gene trn X, cc con c nhn cc gene lin kt gii tnh ch t cc c th m, v vy p th h con bng vi pc th h trc; cc con ci nhn cc gene lin kt gii tnh ng u t c hai b m, v vy pc th h con bng trung bnh cng ca p v pc th h trc. Nu dng du phy trn u ch tn s allele th h con, ta c: p = pc pc = (pc + p) T y xc nh c mc chnh lch hay l hiu s gia cc tn s allele ca hai gii: pc p = (p + pc) pc = (pc p) Ngha l, hiu s ca cc tn s allele gia hai gii th h con bng mt na hiu s ca cc tn s allele gia hai gii th h b m ca n, nhng ngc du. Nh vy, s phn b cc allele gia hai gii c s giao ng theo quy lut sau: C sau mt th h, mc chnh lch gim i mt na v nh th qun th tin dn n trng thi cn bng cho n khi cc tn s gene hai gii l cn bng nhau, ngha l pc = p = p .

V d: Theo kt qu mt mu nghin cu trn mo Lun n (Searle, 1949; trong Falconer 1989) cho thy trong s 338 mo ci c 277 con lng en (BB), 54 con th khm (BO) v 7 con lng da cam (OO), v trong s 353 mo c c 311 en (B) v 42 da cam (O). Tnh trng ny tun theo quy lut di truyn kin kt vi gii tnh nh cp trc y. kim tra xem qun th c trng thi cn bng hay khng, trc tin ta hy xem liu c bng chng no v s giao phi ngu nhin? Php th u tin l xem tn s allele hai gii c ging nhau khng. Tnh ton c th cho thy cc tn s gene hai gii khc nhau khng ng k. - gii ci: f(B) = pc = (2277 ) + 54/( 2338 ) = 0,8994 f(O) = qc = (27 ) + 54/( 2338 ) = 0,1006 - gii c: p = 311/353 = 0,881 T tn s cc allele gii ci, ta tnh c s c th k vng ca mi kiu gene gii ny nh sau:
S c th Kiu gene Tng
BB 277 273,2 P = 0,04
BO 54 61,2
OO 7 3,4 338 338
Quan st K vng 2(1) = 4,6
Kt qu cho thy cc s liu quan st khng ph hp lm vi s k vng m ch yu l cc s liu thp (kiu BO v OO). Nu vy th s khng nht qun c th l do giao phi ngu nhin, nhng cng c th do th hiu ca con ngi thin v cc mu sc lm sai lch mu, khng i din c cho qun th. Qua s phn tch ny cng vi s sai khc cht t v tn s gene gia hai gii ni trn, chng ta chng c l do g ngh rng qun th ny khng trng thi cn bng.
IV. Ni phi (inbreeding)

Bn cnh cc qun th giao phi ngu nhin xt, cn c cc ngoi l i vi gi nh ny mt s loi, chng hn nh cc thc vt hoc ng vt khng xng sng t th tinh. Ni phi l s giao phi khng ngu nhin (nonrandom mating) xy ra gia cc c th c quan h h hng gn; n c tm quan trng c bit ngi, bi v nhiu ngi mang cc bnh di truyn ln sinh ra t s kt hn h hng. Hn na, ni phi c s dng trong chn ging thc vt v c ng vt to ra cc dng mang nhng c tnh mong mun no . S giao phi khng ngu nhin i vi cc kiu gene xy ra trong cc

qun th, trong cc c th giao phi hoc l c quan h h hng gn hn hoc l t gn hn so vi k vng giao phi ngu nhin t qun th. Kt qu ca hai kiu giao phi ny c gi tng ng l ni phi (inbreeding) v ngoi phi (outbreeding). Mc d c hai kiu giao phi ny t n khng lm thay i tn s allele, nhng u lm thay i tn s cc kiu gene ( mi locus trong b gene). * Trong mt qun th ni phi, tn s ca cc th ng hp tng ln v tn s ca cc th d hp gim xung so vi cc t l giao phi ngu nhin. * Trong mt qun th ngoi phi, tnh hnh xy ra ngc li, vi tn s cc th d hp tng v tn s cc th ng hp gim so vi cc t l giao phi ngu nhin . 1. T th tinh (self-fertilization) Kiu cc oan nht ca ni phi l s t th tinh, trong ht phn v non (hay tinh trng v trng) c sinh ra trn cng mt c th. Hnh thc sinh sn ny ph bin mt s nhm thc vt. Trong trng hp t th tinh hon ton, mt qun th c phn thnh nhiu dng ni phi mau chng tr nn ng hp cao . l trng hp cc dng u thun chng c s dng bi Mendel.
Bng 12.6 S bin i tn s kiu gene trong mt qun th khi u vi ch nhng th d hp t th tinh hon ton qua nhiu th h
Tn s kiu gene Th h Tn s allele H s ni phi
0 1 2 ... n
AA 0 1/4 3/8
Aa 1 1/2 1/4
aa 0 1/4 3/8
a 0,5 0,5 0,5
(F)_______ 0 1/2 3/4
1 1/ 2n 2
1/2
1/2 0
1 1/ 2n 2
1/2
0,5 0,5
11/2n 1
minh ha cho iu ny, ta gi s th h b m c ba kiu gene AA, Aa v aa vi tn s tng ng l P, H v Q. Khi s t th phn l hon ton, cc kiu gene AA v aa sinh ra i con tng ng ton l AA v aa; cn kiu gene Aa theo quy lut phn ly Mendel s cho i con gm mt na l Aa v na kia phn ng u cho hai kiu ng hp, AA v aa. Khi tn s ca cc kiu gene AA, Aa v aa th h con tng ng l: (P + H), (H) v (Q + H). Nh vy, sau mt th h t th tinh hon

ton, tn s th d hp gim i mt na so vi b m, trong khi tn s ca hai kiu ng hp tng ln. hiu r s ni phi lm thay i cc t l kiu gene ra sao, ta hy xt qun th ban u gm ch nhng th d hp Aa khi t th tinh hon ton qua nhiu th h (bng 12.6). V c sau mi th h mc d hp t li gim i mt na so vi th h trc n, nn th h th n mc d hp bng ()n ca tr s ban u: Hn = (n)H0, trong H0 v Hn l t l th d hp th h ban u v th h th n. V t l mi kiu ng hp l (1n)/2. Khi n tin n v hn th thnh phn d hp b trit tiu v ch cn li hai thnh phn ng hp vi tn s l . Lc ny, tn s mi kiu gene bng tn s allele. Mt im quan trng ca ni phi l ch, mc d cc tn s kiu gene c th b thay i nhiu, nhng cc tn s allele vn c gi nguyn khng i qua cc th h. Bn c th t chng minh iu ny? 2. H s ni phi (inbreeding coefficient ) m t hiu qu ca ni phi ln cc tn s kiu gene ni chung, ta s dng php o gi l h s ni phi (F). Tr s ny c nh ngha l xc xut m hai allele ti mt locus trong mt c th l ging nhau v ngun gc (cc allele c coi l ging nhau v ngun gc khi hai allele trong mt c th lng bi bt ngun t mt allele c th ca t tin). Tnh cht ca h s ni phi (F): + Tr s F chy t 0 dn 1 (xem ct cui bng 12.6). + F = 1 khi tt c cc kiu gene trong qun th l ng hp cha cc allele ging nhau v ngun gc. + F = 0 khi khng c cc allele ging nhau v ngun gc. + Trong mt qun th ngu phi c kch thc ln, F c coi l gn bng 0, bi v bt k s ni phi no cng c th xy ra gia cc c th h hng rt xa v v vy s c tc dng nh ln h s ni phi . Gi s rng qun th gm ba kiu gene AA, Aa v aa c phn tch thnh mt t l ni phi (F) v mt t l ngu phi (1 - F). Trong qun th ni phi, tn s ca AA, Aa, v aa tng ng l p , 0, v q. y l t l ca cc dng c k vng i vi mi kiu gene, nu nh s t th tinh hon ton din ra lin tc. Bng cch cng cc t l ni phi v ngu phi vi nhau v s dng mi quan h q = 1 p, lc tn s cc kiu gene tr thnh nh sau (xem bng 12.7): P = p2 + Fpq H = 2pq 2Fpq

Q = q2 + Fpq Trong mi phng trnh trn, s hng u l t l H-W ca cc kiu gene v s hng sau l lch so vi tr s . Lu rng cc c th ng hp, v d AA, c th hoc l do hai allele ging nhau v ngun gc, ngha l bt ngun t cng mt allele t tin (s hng Fpq) hoc l do hai allele ging nhau v loi sinh ra qua ngu phi (s hng p2). ln ca h s ni phi phn nh lch ca cc kiu gene so vi cc t l H-W; ngha l, lc F = 0 th cc hp t t t l H-W, v khi F > 0 do c ni phi, th xy ra s gim thiu cc th d hp v di tha cc th ng hp.
Bng 12.7 Tn s ca cc kiu gene khc nhau khi trong qun th xy ra c ni phi ln ngu phi
Kiu gene Ni phi (F) Ngu phi (1 F) Tng
AA Aa aa
Fp Fq F
(1 F)p2 (1 F)2pq (1 F)q2 1F
Fp + (1 F )p2 = p2 + Fpq (1 F)2pq = 2pq 2Fpq Fq + (1 F)q2 = q2 + Fpq 1 1
3. Tnh ton h s ni phi C hai cch c tnh h s ni phi, l da vo cc tn s kiu gene hoc l da vo cc ph h. Vi phng php th nht, ta c tnh h s ni phi trong mt qun th t nhin bng cch s dng biu thc v tn s cc th d hp cho trn. Qua ta c th tm ra biu thc cho F nh sau: H = 2pq 2Fpq = (1 F)2pq 1 F = H/2pq Suy ra F = 1 (H/2pq) T phng trnh trn cho thy h s ni phi (F) l mt hm ca t s gia mc d hp t quan st c (H) v mc d hp t k vng (2pq). Trng hp c ni phi, H nh hn 2pq, v vy F > 0. Nu nh khng c th d hp no c (H = 0), th h s ni phi bng 1. Nhu loi thc vt c h thng giao phi bao gm c t th phn v giao phn t do vi cc c th khc. Nu nh t l t th phn cao, th hu nh tt c cc c th trong qun th l cc th ng hp. V d, mt qun th thc vt gm ba kiu gene AA, Aa v aa vi cc tn s tng ng l P = 0,70, H = 0,04 v Q = 0,26. Ta c th c tnh h s ni phi nh sau : Trc tin, tnh c cc tn s allele A v a (p v q ):

p = 0,70 + (0,04) = 0,72 v q = 1 p = 0,28 Vy h s ni phi F = 1 ( 0,04/2 0,72 0,28 ) = 0,901 Tr s F y rt cao, gi rng hu ht qun th ny sinh sn bng t th phn v ch mt s rt nh l tp giao. Phng php th hai thu nhn h s ni phi cho i con l t mt ph h trong c xy ra s giao phi cn huyt (consanguineous mating). Trong trng hp ny ta s dng mt ph h tnh xc xut ca cc t hp cha cc allele ging nhau v ngun gc i con. V d, ta hy tnh h s ni phi cho mt i con ca hai anh ch em bn ng huyt (half-sibs), tc cc c th sinh ra t cng mt b (hoc m). Hnh 12.1a cho ph h v kiu giao phi ny, trong X v Y l hai anh em c cng m nhng khc cha. Ngi m ca X v Y c biu th l t tin chung (CA = common ancestor). Cn hai ngi cha khng gp phn vo h s ni phi c biu din bng cc hnh vung trng. hnh 12.1b, cng mt ph h nh th nhng biu din theo mt cch khc, b qua cc k hiu cha m cn cc du qu trm biu th cho tt c cc c th, v gii tnh khng quan trng trong vic xc nh h s ni phi y. Cc mi tn trn hnh v ch hng truyn t b m n con ci. I II III
(a) CA
CA Y X
(b)
X Z
Y Z
Hnh 12.1 Ph h minh ha s kt hn gia hai anh em bn ng huyt, X v Y. (a) vi tt c cc c th; (b) khng c b. y CA = t tin chung, v ng k i ch s giao phi cn huyt.
Gi s ngi m (CA) c kiu gene l Aa. tnh h s ni phi, ta cn phi bit xc sut m a chu ca b, Z, c kiu gene AA hoc aa, l ging nhau v ngun gc i vi mt trong hai allele ca b. Trc tin ta xt Z l AA, ch c th xy ra nu nh mi bn X v Y u ng gp vo Z mt giao t cha A. Xc sut ca allele A trong X l xc sut m mt allele A n t CA, hay . V xc sut truyn t allele A t X sang Z cng l , nn xc sut kt hp ca hai s kin ny l = (qui tc nhn xc sut). Tng t, xc sut Z nhn c allele A t Y l . V vy xc sut ca mt a con AA nhn c allele A t mi bn X v Y

l = 1/16 hay 0,0625. Bng phng php ny ta tnh c xc sut ca mt a con c kiu gene aa l 1/16. Nh vy xc sut ton b cc t hp c cha cc allele ging nhau v ngun gc Z lc l 1/16 + 1/16 = 1/8 hay 0,125 (qui tc cng xc sut ). n gin, trong tnh ton h s ni phi t mt ph h ngi ta xut mt phng php gi l k thut m chui (chain-counting technique). Mt chui i vi mt t tin chung cho trc bt u vi mt b m ca c th ni phi, ngc tr ln ph h cho n t tin chung, v tr li vi b m . V d, t hnh 12.1 ta lp c chui n gin X-CAY. S c th trong chui (n) c dng tnh h s ni phi trong cng thc sau y: F = (1/2)n. Vi v d trn, h s ni phi l (1/2)3 = 0,125.
V. Cc nhn t tc ng ln thnh phn di truyn qun th

Trong t nhin, thnh phn di truyn ca cc qun th ni chung l khng n nh nh nguyn l Hardy-Weinberg vch ra, m lun lun b bin ng di nh hng ca nhiu nhn t khc nhau. Ln u tin, vn ny c Charles Darwin nu ln trong tc phm Ngun gc cc loi bng con ng chn lc t nhin (On the Origin of Species by Means of Natural Selection) nm 1859. Theo quan nim hin nay, c bn nhn t c bn lm thay i tn s cc allele ca cc qun th v xc nh qu trnh tin ha, l: t bin, bin ng di truyn ngu nhin, di-nhp c v chn lc t nhin. V chi tit, c th xem thm trong Bi ging Di truyn hc qun th (Hong Trng Phn 2003); y chng ta ch tm hiu s lc vai tr v hiu qu ca mi nhn t. 1. t bin t bin (mutation) c nhiu loi khc nhau nh c trnh by cc chng 3 v 8; y chng ta ch cp n vai tr, tnh cht v p lc ca cc t bin gene t pht i vi qu trnh tin ha v chn lc. Phn ln t bin mi xut hin c hi cho c th t bin l ngun cung cp ch yu cc bin d di truyn mi trong mt qun th-loi, v vy n c xem l mt qu trnh quan trng c bit trong di truyn hc qun th. Ni chung, phn ln t bin mi xut hin l c hi, mt s t bin l trung tnh v ch mt s t l c li cho bn thn sinh vt. Theo thuyt trung tnh (Kimura 1983), i a s cc bin i tin ha cp phn t c gy nn khng phi bng chn lc Darwin m bng s c nh ngu nhin cc th t bin trung tnh hoc hu nh trung tnh v mt chn lc; p lc t bin v bin ng di truyn ngu nhin chim u th trong s bin i tin ha cp phn t. xt xem hiu qu ca t bin ln s bin i di truyn trong mt

qun th, ta xt hai allele A (kiu di) v a (gy hi) vi tn s ban u tng ng l p v q; gi u l t l t bin thun t A thnh a cho mt giao t mi th h, v v l t l t bin nghch t a thnh A. Cc allele A do t bin thun thnh a lm tng tn s ca allele a ln mt lng l up, trong khi tn s alllele a do t bin nghch c th b gim i mt lng l vq. Nh vy, nhn ton cc th sau mi th h s bin i trong tn s ca allele a (q) do t bin l: q = up vq Tr s dng cc i cho s bin i ny l u, khi p = 1 v q = 0 (ngha l tt c cc allele u l kiu di). Tr s m cc i l v, khi p = 0 v q = 1. Tuy nhin do cc t l t bin u v v ni chung l nh, nn s bin i c k vng do t bin cng rt nh. Chng hn, nu ta cho u = 10-5, v = 10-6 v q = 0,0, lc : q = (0,00001)(1,0) (0,000001)(0,0) = 0,00001 Mc d t bin ch gy mt hiu qu nh trong tn s allele mi th h, nhng n li c tm quan trng cn bn trong vic xc nh mc gy ra cc bnh di truyn him. Trn thc t, s cn bng gia t bin (lm tng tn s ca allele bnh) v chn lc (lm gim tn s ca allele bnh) c th l gii mc quan st c ca cc bnh nh bch tng chng hn. Ngoi ra, t bin cng vi s bin ng di truyn ngu nhin cho php gii thch hp l cho s lng ln cc bin i phn t quan st c gn y nhiu loi (xem Kimura 1983). 2. Bin ng di truyn ngu nhin Bin ng di truyn ngu nhin (genetic random drift), hay ni gn l bin ng di truyn, l nhng s bin i ngu nhin v hng v tn s allele trong tt c cc qun th, nhng c bit l cc qun th nh. Bin ng di truyn l mt qu trnh thun ty ngu nhin, mang tnh xc sut v t l nghch vi kch thc qun th. Trong mt qun th ln, thng thng bin ng di truyn ch gy ra mt s thay i ngu nhin nh. Nhng trong cc qun th nh, n c th gy ra nhng s bin ng ln v tn s allele qua cc th h khc nhau. Chnh hin tng ny l nguyn nhn to nn s khc bit a dng v mt di truyn cc qun th nh v dn dn dn ti s cch ly sinh sn trong qu trnh tin ha ca loi. V s bin ng di truyn c th l nguyn nhn lm cho mc d hp t thp quan st c mt s loi c nguy c b dit vong. 3. Dng gene hay s di nhp c Di-nhp c (migration) hay dng gene (gene flow) l s di chuyn ca cc c th t mt qun th ny sang mt qun th khc, ko theo vic a

vo cc allele nhp c mi thng qua s giao phi v sinh sn sau . Nh vy, dng gene khng lm thay i cc tn s allele ca c loi, nhng c th lm bin i cc b (tn s allele so vi nguyn l H-W) khi tn s cc allele ca nhng c th di c ti l khc vi ca cc c th c tr ti ch. Gi s cc c th t cc qun th xung quanh di c ti mt qun th a phng ta nghin cu vi tc m mi th h v giao phi vi cc c th c tr . V cng gi s rng tn s allele A ca qun th ngun gene nhp c l P v ca qun th nghin cu l po. Khi , th h th nht sau nhp c, tn s allele A ca qun th nghin cu s l: p1 = (1 m)po + mP = po m(po P) S bin i p v tn s allele sau mt th h l: p = p1 po Thay tr s p1 thu c trn, ta c: p = m(po P) iu c ngha l, t l cc c th di c cng ln v s chnh lch gia hai tn s allele cng ln, th i lng p cng ln. Lu rng p = 0 ch khi hoc m = 0 hoc (po P) = 0. Nh vy, tr phi s di c dng li (m = 0) cn th tn s allele s tip tc bin i cho n khi n tr nn ging nhau gia qun th a phng v qun th ph cn (po P = 0). Sau th h th nht, hiu s v tn s allele gia hai qun th trn l: p1 P = po m(po P) P = (1 m)(po P) Tng t, sau th h th hai, hiu s v tn s allele s l: p2 P = (1 m)2.(po P) V sau n th h di c, ta c: pn P = (1 m)n.(po P) Cng thc ny cho php ta tnh ton hiu qu ca n th h di c tc m no , nu nh bit c tn s cc allele khi u (po v P): pn = (1 m)n.(po P) + P Hoc nu nh bit c tn s cc allele khi u (po v P), tn s allele ca qun th nghin cu ti mt thi im no (pn) cng nh s th h n, ta c th tnh c tc dng gene (m). V d: M (USA), nhng ngi c ngun gc hn chng da trng Capca (Caucasian) v da en Chu Phi (African) c coi l thuc qun th ngi da en. S pha tp v chng tc c th xem nh l mt qu trnh ca dng gene t qun th Capca sang qun th da en. Tn s ca allele Ro locus xc nh cc nhm mu rhesus l P = 0,028 cc qun th Capca nc M. Trong s cc qun th Chu Phi m t cc t tin ca

ngi M da en di c n, tn s allele Ro l 0,630. T tin Chu Phi ca nhng ngi M da en n nc M cch y khong 300 nm hay khong 10 th h; ngha l n = 10. Tn s allele Ro trong s nhng ngi M hin gi l pn = 0,446. Bng cch bin i li phng trnh trn, ta c: (1 m)n = (pn P) : (po P) Thay cc tr s cho, ta c: (1 m)10 = (0,446 0,028) : (0,630 0,028) = 0,694 1m=
10
0,694 = 0,964
Suy ra: m = 0,036 Nh vy, dng gene chuyn t nhng ngi M Capca vo trong qun th ngi M da en xy ra tc tng ng vi tr s trung bnh l 3,6% mi th h. Nhng tnh ton tng t bng cch s dng cc tn s allele nhiu locus khc cho cc kt qu hi khc mt cht. Hn na, mc pha tp hn chng c th khc nhau cc vng khc nhau ca nc M; nhng r rng l s trao i gene ng k xy ra (Ayala v Kiger 1980, tr.644; Hartl et al 1988, tr.214). 4. Chn lc t nhin C ba qu trnh gy bin i tn s gene xt trn y u c mt im chung l khng mt qu trnh no nh hng i vi s thch nghi (adaptation). Theo nhn nh ca mt s tc gi (Mayer 1974; Ayala v Kiger 1980), cc qu trnh ny l ngu nhin i vi s thch nghi, do t thn chng s ph hoi t chc v cc c tnh thch nghi ca sinh vt. Ch c chn lc t nhin (natural selection) mi l qu trnh thc y s thch nghi v hn ch cc hiu qu ph hoi t chc ca cc qu trnh khc. Trong ngha , chn lc t nhin l qu trnh tin ha khc lit nht, bi v ch c n mi c th gii thch c bn cht thch nghi, tnh a dng (diversity) v c t chc cao ca cc sinh vt. tng v chn lc t nhin nh l qu trnh nn tng, l ng lc ca s bin i tin ha do Charles Darwin v Alfred Russel Wallace c lp a ra nm 1858. L lun tin ha bng chn lc t nhin c pht trin y , vi chng c ng h xc ng, trong cun Ngun gc cc loi (The Origin of Species) do Darwin xut bn nm 1859. Theo Hartl et al (1988, 1997), trn quan im thuyt tng hp hin i, c th hnh dung chn lc t nhin xy ra da trn ba im chnh: (1) mi sinh vt, i con c sinh ra nhiu hn s sng st v sinh sn; (2) Cc c th khc nhau v kh nng sng st v sinh sn v phn ln nhng

khc bit ny l do kiu gene; (3) Trong mi th h, cc kiu gene sng st s sinh sn nhiu hn v quyt nh s phn b li cc kiu gene th h sau. Hu qu l, cc allele tng cng s sng st v sinh sn s gia tng tn s t th h ny sang th h khc, v qun th s ngy cng sng st v sinh sn tt hn vi mi trng ca n. Trn quan im , chn lc t nhin c nh ngha l s sng st v sinh sn bit ha ca cc kiu gene. hiu c cc tc dng ca chn lc ln bin d di truyn, ta phi xt xem ph hp tng i (relative fitness) ca cc kiu gene khc nhau. N c nhiu thut ng ng ngha nh: ph hp Darwin (Darwinian fitness), gi tr chn lc (selective value), hay gi tr thch nghi (adaptive value). Tu trung, n c th c nh ngha nh l kh nng tng i ca cc kiu gene khc nhau trong vic truyn li cc allele cho nhng th h tng lai (Weaver v Hedrick 1997). Bi v chn lc t nhin tc ng bng s sinh sn bit ha, nn ta c th xem n l s o hiu nng sinh sn ca mt kiu gene. cho tin, cc nh di truyn hc thng t nh tr s ph hp (w) bng 1 cho kiu gene c hiu nng sinh sn cao nht. Mt n v o lin quan l h s chn lc (selection coefficient), c k hiy bng s, v c nh ngha l s = 1 w. H s chn lc o mc gim bt ph hp ca mt kiu gene. Gi s mi th h cc kiu gene AA v Aa u sinh c 100 con, cn th ng hp ln sinh c 80 con; nu ta coi ph hp ca cc c th mang allele tri l 1, th ph hp ca cc th ng hp ln l 0,8. Hiu s ca cc tr s ph hp ny chnh l h s chn lc (s), v trong trng hp ny s = 1 0,8 = 0,2. Nu nh cc kiu gene c kh nng sng st v sinh sn nh nhau th s = 0; nu mt kiu gene no gy cht hoc lm bt th hon ton th s = 1. 4.1. Chn lc v t bin Chn lc c xu hng o thi cc allele c hi ra khi qun th, trong khi t bin c th to ra cc allele c hi mi. Qun th s gi nguyn trng thi phn b cc kiu gene nu nh tn s t bin mi xut hin va ng bng tn s allele b chn lc o thi. Sau y ta th xt s cn bng ny trong trng hp "Chn lc chng li cc ng hp t ln". Gi s A l allele bnh thng v a l allele c hi vi tn s tng ng ca chng l p v q. Khi ph hp hay gi tr thch nghi ca cc kiu gene AA, Aa v aa tng ng l 1: 1: 1-s. Trong trng hp ny tc o thi allele a khi qun th bi chn lc s l sq2. Nu cho rng tc t bin thun (A a) l u, th tc xut hin allele a mi trong qun th l up. V p 1 (do tn s a rt thp) nn c th coi up u. Vi c ch

ngu phi, qun th s trng thi cn bng khi tc xut hin t bin mi bng tc o thi, ngha l u = sq2, hay khi tn s allele ln trong qun th mc q = u / s . Tng t, i vi allele tri, u = sp hay p = u/s. V d: Tn s mc bnh PKU tr s sinh l khang 4 trn 100.000; do q2 = 410-5. Hiu qu sinh sn ca cc bnh nhn khng c cha tr l zero, hay s = 1. Khi u = sq2 = 4 10-5.
4 x10 5 = 6,310-3 v tn s ca cc th d hp l: 2pq 2q = 2(6,310-3) = 1,2610-2 iu c ngha l, trong 100 ngi c khong 1,3 ngi mang allele , mc d c 4 trong 100.000 ngi mc bnh PKU. Tn s ca allele ny c mt trong cc th d hp bng mt na ca 1,2610-2 hay 6,310-3; v tn s ca allele cc th ng hp l 4 10-5. Do vy cc allele PKU c mt trong cc th d hp nhiu hn 6,310-3 / 4 10-5 = 158 ln so vi cc th ng hp. Nh ni t u, cc allele him tn ti trong qun th hu ht cc th d hp. 4.2. u th d hp t u th d hp t hay siu tri (overdominance) l trng hp chn lc u i ng h cc th d hp nhiu hn c hai dng ng hp t. Khi chn lc khng loi thi allele no. mi th h, cc th d hp s sinh sn mnh cho nhiu con chu hn cc th ng hp v chn lc s gi li c hai allele cho n khi qun th to c trng thi cn bng, vi cc tn s allele khng thay i. Mt v d ni bt v hin tng siu tri trong cc qun th ngi l bnh thiu mu hng cu hnh lim, mt bnh ph bin chu Phi v chu . Bnh ny c lin quan n mt dng st rt do k sinh trng ph bin gy ra l Plasmodium falciparum. Allele HbS gy cht trc tui trng thnh nhng ngi ng hp t HbSHbS. Tn s allele ny c th cao hn 10% cc vng c st rt ni trn, bi v cc th d hp HbAHbS khng c s nhim st rt, trong khi cc th ng hp HbAHbA th khng c kh nng .
Tn s allele ny trong cc qun th ngi l q =
Cu hi v Bi tp
1. (a) nh ngha cc khi nim sau: di truyn hc qun th, qun th, vn gene, tn s allele v tn s kiu gene. (b) c trng di truyn ca cc qun th giao phi v ni phi l g? 2. Ti sao ni khi nim cn bn v cng c thit yu ca di truyn hc

qun th l tn s allele ch khng phi tn s kiu gene? Hy thit lp cc cng thc tnh tn s allele khc nhau v cho v d. 3. (a) Nu ni dung v cc iu kin nghim ng ca nguyn l Hardy-Weinberg. T hy chng minh v nu cc h qu v ng dng ca n. (b) Th no l trng thi cn bng di truyn ca qun th? Cc phng php kho st trng thi cn bng di truyn ca qun th? 4. Nguyn l Hardy-Weinberg c m rng cho cc trng hp a allele, tn s allele sai khc gia hai gii tnh v cc gene lin kt vi gii tnh nh th no? 5. Hy tnh tn s cc allele v kho st trng thi cn bng theo cc cch khc nhau i vi h nhm mu M-N cc qun th ngi sau y (s liu t F.J. Ayala v J.A. Kiger 1980):
Qun th M MN N Tng
Th dn c 22 216 492 730 B lc da Navaho 305 52 4 361 Ngi M gc Capca 1787 3039 1303 6129 6. Hy tnh tn s allele trong cc trng hp sau: a) Cc th ng hp ln nhiu gp hai ln cc th d hp. b) Cc th ng hp ln nhiu gp su ln cc th d hp. 7. Mu sc v c sn chu u c kim sot bi ba allele mt locus n: CB (nu), CP (hng) v CY (vng). Allele mu nu l tri so vi hng v vng; mu hng tri so vi vng; mu vng l ln hon ton. Trong mt qun th c sn cc mu sc c phn b nh sau: 472 nu, 462 hng, 66 vng. Nu nh qun th ny dng cn bng, tn s cc allele s ra sao? 8. Chng m mu do mt allele ln m lin kt trn X gy ra. C mi ngi c mt ngi nam mc chng m mu. Hi: (a) T l n gii l bao nhiu? (b) Nu mt na s tr sinh ra mi gii tnh b m mu, th t l cc cuc hn nhn gy ra hu qu l bao nhiu? (c) Nu nh tt c tr sinh ra u bnh thng, th t l cc cuc hn nhn l nh th no? 9. Chng minh rng khi mt qun th tin ti trng thi cn bng i vi mt gene lin kt gii tnh, th: (a) S dao ng v tn s allele gia hai gii thu hp dn sau mi th h theo quy lut sau: (pc p) = (pc p) , trong pc v p l cc tn s gene tng ng ca hai gii ci v c i b m, cn pc v p l cc tn s tng ng i con ca n. (b) Tn s gene ca c qun th ( p ) l khng i qua cc th h v bng pc + p.

10. (a) mt qun th sinh vt, tn s ca cc th ng hp tri, d hp v ng hp ln tng ng l 0,67; 0,06 v 0,27. H s ni phi l bao nhiu? (b) Nu nh trong mt qun th vi hai allele mt locus nhim sc th thng c p = 0,8, q = 0,2, v tn s ca cc th d hp l 0,20, th h s ni phi l bao nhiu?
Ti liu Tham kho

Ting Vit Trn B Honh. 1988. Hc thuyt tin ha. NXB Gio Dc, H Ni. Kimura M. 1983. Thuyt tin ha phn t trung tnh (Bn dch ca Hong Trng Phn). NXB Thun Ha, Hu -1993. Mayer E. 1974. Qun th, loi v tin ha. (Bn dch ca Lng Ngc Ton, Hong c Nhun, Nguyn c Khm v Nhuyn Vn Tho). NXB Khoa hc v K thut, H Ni, 1981. Hong Trng Phn. 2001. H qu ca mt s nh lut di truyn v ng dng ca chng trong ging dy di truyn hc. Trong: K yu Hi ngh Khoa hc Khoa Sinh-KTNN, Trng HSP H Ni, tr.5-8. Hong Trng Phn. 2003. Di truyn hc qun th (Bi ging lu hnh ni b). Trng HSP Hu. Ting Anh Ayala FJ, Kiger JA. 1980. Modern Genetics. Benjamin/Cummings Publishing Company, Inc, Menlo Park, CA. Crow JF. 1986. Basic Concepts in Population, Quantitative, and Evolutionary Genetics. WH Freeman and Co., New York. Crow JF. 1993. Mutation, mean fitness, and genetic load. In: Oxford in Evolutionary Biology, Volume 9 (Futuyma D., Antonovics J. eds), pp 3-42, Oxford University Press, Inc., Oxford -New York... Falconer DS, Mackay TFC. 1996. Introduction to Quantitative Genetics. 4th ed., Longman Group, Harlow. Hartl DL, Freifelder D, Snyder LA. 1988. Basic Genetics. Jones and Bartlett Publishers, Inc., Boston - Portola Valley. Hartl DL, Clark AG. 1997. Principles of Population Genetics. 3rd ed., Sinauer Associates Inc., Sunderland, Massachusets. ISBN: 0-87893-306-9. Holman DJ. 2003. Human Population Genetics BIO A 482 Autumn 2003). <http://faculty.washington.edu/djholman/bioa482/> Ridley M. 1993. Evolution. Blackwell Scientific Publishing, Boston.

Standfield WD. 2002. Genetics, 4th ed. Schaum's Outline Series. McGraw-Hill Inc., Toronto. Suzuki DT, Griffiths AJF, Miller JH, Lewontin RC. 1989. An Introduction to Genetic Analysis. 4th ed., W-H Freeman and Company, New York. Yablokov V.A. 1986. Population Biology: Progress and Problems on Natural Populations. Mir Publishers, Moscow. Weaver RF, Hedrick PW. 1997. Genetics. 3rd ed., McGraw-Hill Companies, Inc./ Wm.C.Browm Publishers, Dubuque, IA. Wrights 1988. Surfaces of selective value revisited. The American Naturalist 131: 115-123. Mt s trang web http://wbar.uta.edu/jands/jands.htm http://science.kennesaw.edu/~rmatson/Biol%203380/3380species.html http://www.users.nac.net/jmele/ECO.HW.html http://nitro.biosci.arizona.edu/courses/EEB182/Lecture04/lect4.html

Giao Trinh Di Truyen Hoc - Hoang Trong Phan, Truong Thi Bich Phuong, Tran Quoc Dung

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Chng 1: C s ca Di truyn hc Mendel

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Chng 3: C s T bo ca s Sinh sn, Di truyn v Bin d

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Hong Trng Phn 103

Chng 5: Bn cht Ho hc v Ti bn ca Vt cht Di truyn

147 147 148 149 150 150

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Chng 6: Gene v Qu trnh Sinh tng hp Protein

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Chng 7: S iu ho Biu hin ca Gene

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Trng Th Bch Phng 223

223 223 228 228 232 237 237 239

Chng 9: S Di truyn T bo cht

Trng Th Bch Phng 245

Chng 10: i cng v Cng ngh DNA Ti t hp

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267 269 269 271 274

Chng 11: Di truyn hc Ngi

Chng 12: Di truyn hc Qun th

I. Cc khi nim c bn ca di truyn hc qun th 1. Qun th (population) 2. Cc h thng giao phi

296 296 296

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II. Lc s pht trin ca di truyn hc

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Hnh 4 Beadle, Tatum, Jacob v Monod (t tri sang)

Hnh 6 R.Franklin (tri) v M.Wilkins

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III. i tng v cc lnh vc nghin cu ca di truyn hc

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IV. Cc phng php nghin cu ca di truyn hc

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