T CHC PHN T

CA NHIM SC TH
Nhm 3:
Trng Xun Lc
Tn N Hong Phng
Phm Th Thanh Hng
Nguyn Th Tuyt
I. KHAI NIM V NHIM SC TH (NST)
-NST l c s di truyn cp t bo.
- L cu truc nm trong nhn t bo, co kha nng bt
mu c trng vi cac phm nhum kim tinh.
- Mi loi sinh vt co b NST c trng v hinh dng,
kich thc, s lng v c duy tri n nh qua cac
th h.
Hinh dng NST
II. CU TRUC NST CUA PROKARYOTE
- Sinh vt nhn s (Prokaryote) l nhom sinh vt m t
bo khng co mng nhn, do o nhn khng c hnh
dng c nh vng nhn.
- Khi nhum mu t bo bng thuc nhum Feulgen,
vng nhn hin mu tm.
- Nhim sc th cha 1 si ADN dng vng xon kp (
X khunStreptomyces c th gp nhim sc th dng
thng).
- NST ca prokaryote c kch thc thay i t
160.000 cp base vi khun Candidatus Carsonella
ruddii n 12.200.000 cp base vi khun sng
trong t Sorangium Cellulosum.
- Mt s vi khun nh Borrellia (Xon khun
gy bnh Lyme) c cha nhim sc th tuyn tnh
(linear chromosome).
Linear choromosome
loi Borrelia burgdorferi NEU2011
- Vi khun Escherichia coli l i tng c s dung
rng rai trong cac nghin cu di truyn phn t.
Vi khun Escherichia coli
III. CU TRUC NST CUA EUKARYOTE
1. Cu truc hin vi cua NST
-Mi NST gm 2 cromatic gn
vi nhau tm ng, mi
cromatic cha 1 phn t ADN.
- Mi NST in hinh u cha
cac trinh t nucleotit c bit
c goi l tm ng v cac
trinh t nucleotit 2 u cung
ca NST c goi l u mut
v trinh t khi u nhn i
ADN.
-Cac NST thng khac nhau v v tri tm ng
(centromere), l ni cac si thoi v sc bam vo
trong qua trinh phn bo.
-Cac tm ng co ban cht l ADN v kinetochore
bam quanh no co ban cht protein.
S kt hp ca 2 thnh phn ny to thnh 1 cu
truc ng hoc tham gia vo cac khia cnh thit yu
ca vic di chuyn ca NST v cac t bo con trong
qua trinh phn bo.
-Mi NST thng chi co mt eo tht ln cha mt tm
ng goi l eo s cp (primary constriction)
-NST thnh 3 kiu chinh:
+ NST tm gia
+ NST tm u
+ NST tm mut
- Ngoi eo s cp ra thi trn mt s NST cu th con co
eo th cp (secondary constriction), nu eo ny nm
gn u mut v xay ra s tht su se to nn 1 v
tinh (satellite) ca NST ( ngi o l cac NST 13, 14,
15, 21 v 22).
- Eo th cp thng cha cac gen tng hp cac ARN
ribosome, tich tu tm thi v hinh thnh nn t chc
goi l hch nhn.
Cu truc u mut cua NST (telome)
- Trn mi NST co cac u mut vi cu truc c trng
goi l telomere, cac telomere khng cha cac gen,
chung gm nhng trinh t ngn (6-8 bp) lp li ni tip
ca ngn ln v co tinh bao tn cao trong tin hoa.
Vi du: Tetrahymena: (TTGGGG)
n

Caenorhabditis: (CCCTCCC)
n
ngi v cac ng vt co vu cac on lp ni tip
(1.000- 2.000 ln) ca telomere nh sau: 5

-TTAGGG-3


- Cac trinh t u mut kt hp vi nhiu loi protein c
trng to thnh cu truc dng vong v co cac chc nng
quan trong nh: n nh v bao v cac u mut ca NST
v c xem l ng h nguyn phn, gii hn s
phn chia ca cac t bo soma trong khoang 40-50 ln.
Cac tm ng v u mut ca NST eukaryote
c cu truc vi nhng trinh t n gian,
c lp li rt nhiu ln goi l ADN v tinh hay
ADN lp li kiu ni tip.
- Mi NST eukaryote l mt phc hp nucleoprotein bao
gm 1 phn t AND si kep mch thng kt hp vi cac
phn t protein c s goi l histone (giu cac aminoacid
lysine v arginine) (2H2A, 2H2B giu lysin, 2H3 v 2H4
giu Arginin. ), phc hp nh th goi l cht nhim sc
(chromatin).
2. Cu truc siu hin vi
- n v t chc c s ca cac NST eukaryote l cac
nucleosome , mi nucleosome co ng kinh khoang 11
nm, gm 1 khi cu 8 phn t histon,
(H2A+H2B+H3+H4)
2
goi l loi octamer v on ADN co
kich thc 146 bp, qun vong xung quanh no ( ni
chung l 1 on ADN di 160- 200 bp qun 2 vong
quanh octamer).
- Mt phn t H1 bam vo on ADN ni gia 2
nucleosom di khoang 150 bp, gi vng s tng tac
ca ADN vi cac histon loi.
- Bc cu truc u tin ca chromatin c hinh
dung di dng mt chui cac nucleosome, hay si
nucleosome co dy 11nm hay ( 110 A
0
).
3. Bn bc cu truc siu hin vi cua NST
- Bc th 2 ca NST l s cun gp chromatin co
lin quan n s xon li ca si nucleosome
thnh si dy 30nm goi l 1 solenoid. Cac histon H1
tham gia vo s xon li ny bng cach tng tac
vi cac phn t H1 khac.
-Bc th 4 ca NST: cac day euchromatin c sp
xp trong cac khng gian 3 chiu xon cht li to
thnh cac vung goi l heterochromatin trn 1
chromatid vi dy 700nm.
- Bc th 3 ca NST l s cun vong ca si
solenoid thnh 1 cu truc xon hn vi cac vong
c neo dinh vo 1 gia trung tm ( dy 300nm). =>
y chinh l vung thao gian xon ca NST, goi l
euchromatic.
Nh vy, s khac nhau c ban gia cu
truc NST vi khun v eukaryote l:
+ sinh vt nhn s cha co cu truc NST
in hinh, NST ca vi khun thng l 1
phn t ADN dng vong cha 1 si ADN
xon kep ( AND khng co s kin kt vi
protein loi histon) v s lng it.
+ sinh vt nhn thc a co cu truc NST in hinh,
s lng ADN nhiu v dng xon kep, a lin kt
vi protein loi histon.
-ADN l cht trng phn (polymer) sinh hoc, c trong
lng phn t ln, gm nhiu n phn ni vi nhau
thnh cc mch (chui) di.
- n phn to nn cc axit nucleic l bn loi nucleic,
trong ADN l cc n deoxyibonucleotid.
- Mi nucleotid gm 3 thnh phn: nhm phosphat,
ng pentose v mt trong bn loi bazo nito (cc
dn xut purin hoc pyrimidin).
- Mi bazo nito gn vi dezoxiibo c goi l
nucleozit. Khi c mt gc axit photphoric gn vo
nucleozit th se thnh mt nucleotit.
- Cc nucleozit ng vi cc bazo c trong thnh phn
ca chng, c goi l xitidin, timidin, adenozin v
guanozin. Cc nucleotide tng ng c goi l
xitidin, timidin, adenin, guanin.

IV. T CHC TRT T NUCLEOTIDE TRONG ADN
- C 4 loi nucltit l A, T, G, X, chng phn bit nhau
v baz nit nn ngi ta goi tn ca cc nucltit
theo tn ca cc baz nit ( A = Anin, T = Timin, G
= Guanin v X = Xitzin).
-Cc nucltit trn mt mch lin kt vi nhau bng lin
kt phtphoieste to thnh chui plinucltit (lin kt
phosphodieste gia gc ng ca nucleotide ny vi
gc phosphate ca nucleotide tip theo).
- Cc nucltit gia 2 mch lin kt vi nhau bng lin
kt hir theo nguyn tc b sung (A ca mch ny lin
kt vi T ca mch kia bng 2 lin kt hir v ngc li;
G ca mch ny lin kt vi X ca mch kia bng 3 lin
kt hir v ngc li).
-Cc lin kt phtphodieste gia cc nucltit trong
chui plinucltit l cc lin kt bn vng, chi nhng
tc nhn t bin c cng mnh mi c th lm
anh hng ti lin kt ny do o lin kt
phtphodieste gi cho phn t ADN s bn vng nht
nh.
- Ngc li, lin kt hir l lin kt yu nhng ADN
c rt nhiu lin kt hir nn ADN va bn vng va
linh hot, chnh nh tnh linh hot ny m cc enzim
c th sa cha cc sai st v trnh t sp xp cc
nucltit.

-Do cu to ho hoc ca cc nucleobase m lin kt
hydro chi hnh thnh gia 2 loi nucleobase nht nh
l A vi T (qua 2 lin kt hydro) v C vi G (bng 3 lin
kt hydro).
- o thc cht l lin kt gia mt purine v 1
pyrimidine nn khoang cch tng i gia 2 chui
polynucleotide c gi vng. Nguyn tc hnh thnh
lin kt trn c goi l nguyn tc b sung v n ph
bin trn moi loi sinh vt.
-Trong t bo, di tc dung ca mt s protein c
hiu, 2 chui ca phn t ADN c th tch nhau ra (cn
goi l bin tnh ADN) do cc lin kt hydro b ct t.
- Khi o, cc nucleotide trn mi chui c th to thnh
lin kt vi cc nucleotide t do trong mi trng ni
bo.
- Kt qua ca qu trnh ny l to thnh 2 phn t ADN
ging ht nhau t 1 phn t ADN ban u.
- y cng chnh l ngun gc ca c tnh di truyn
ca sinh vt.
- Cc nh khoa hoc c th thc hin qu trnh t nhn
i ny trong ng nghim goi l k thut PCR.
Nu s bt cp gia nucleotide chui gc vi
nucleotide khng tun theo nguyn tc b sung th
s to thnh t bin, ngun gc cua hin tng
bin d.



- Trnh t Nucleotit ca gen
cu trc qui nh trnh t
axit amin ca chui
polipeptit.
Nhm Eukaryote tin ho hn vi c im:
99% genome nm trong nhn TB. Phn cn li nm
trong mt s c quan t (ty th, lp th).
+ B gen t bo cht gm cc ADN si kp, vng.
+ B gen trong nhn cha nhiu NST.
Trong t bo eukaryote, DNA kt hp cht che vi
cc protein l histone.
-Cac DNA eukaryote co c im khac vi DNA
prokaryote.
- Ton b phn t DNA prokaryote u mang thng tin
ma ha cho cac protein trong khi o DNA ca
eukaryote bao gm nhng trinh t ma ha (cac exon)
xen ke vi nhng trinh t khng ma ha (intron).

3. Cc yu t di truyn vn ng
prokaryote v eukaryote
2. S pht hin cc yu t
di truyn vn ng
1. Khi nim
4. Cc yu t di truyn vn ng ngi
1. Khi nim

- Yu t di truyn vn ng (transposatic genetic
elements) hay cn goi l gen nhay l on nucleotit c
kha nng di chuyn v tr ngay trong mt th nhim sc
hoc t th nhim sc ny sang th nhim sc khc.
- Gen nhay c vai tr trong s ti t hp li h gen.

2. S pht hin yu t di truyn vn ng
- Barbara McKlintock pht hin v
nghin cu v cc yu t di truyn
vn ng trn cy ng t nhng nm
40 ca th ki XX.
- Vo nm 1983 b nhn c giai
thng Nobel.
- Gery Fink v cs. l nhng ngi u tin s dung
nm men nghin cu iu ho hot tnh gene
eukaryote.
- Vo nm 1985, J.Bocke v G. Fink a chng
minh, yu t Ty1, ging vi retrovirus.

3. Cc yu t di truyn vn ng
prokaryote v eukaryote


Trinh t on xen
Gen nhay
C ch ca s chuyn v
Yu t TnA

Yu t
di truyn vn ng
prokaryote
Cc retrotransposon
DNA transposon
Yu t
di truyn vn ng
eukaryote
3.1.Yu t di truyn vn ng prokaryote
a. Trnh t on xen
-Trnh t on xen (insertion sequence) l mt on
ADN 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.
- Yu t IS khi xen vo gia gen c th lm gin on
trnh t m ha v lm bt hot s biu hin ca gene.

Mt vi trinh t v kich thc ca on xen:
Trinh t
xen vo
S ban sao xen vo
trong E.coli
Chiu
di (bp)
on lp li ao
ngc (bp)
IS1 5 - 8 ban sao trn
nhim sc th
768 18 - 23
IS2 5 ban sao trn NST
1 ban sao trn plasmid
1327 32 - 41
IS3 1400 32 - 38
IS4 1 - 2 ban sao trn
nhim sc th
1400 16 - 18
IS5 Cha bit 1250 Ngn
5 ban sao trn NST
2 ban sao trn plasmid
- Tt ca cc yu t IS u cha on ADN m ha cho
protein, c goi 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 gen ny nm gia 2 on lp li ao ngc
(inverted repeat - IR) ngn.
- V du: S ti t hp ca plasmid v nhim sc th ca
E.coli to ra nhng chng Hfr qua trao i cho n gia
yu t IS trn plasmid v yu t IS trn nhim sc th.
b. Gene nhy cua prokaryote
-Cc yu t IS ring l khng chi c kha nng t di
chuyn m khi hai yu t ny nm gn nhau th
chng c th vn ng nh mt n v hon chinh v
mang theo cc gene nm gia chng.
- Cu trc phc tp ny c goi l transposon.
- C hai kiu transposon vi khun:
+ Transposon hn hp (composite transposon)
+ Transsposon n gian (simple transposon)

Hnh 1. c trng v cu trc ca transposon hn hp
(composite transposon) v transsposon n gian (simple
transposon)

c. C ch cua s chuyn v
- S chuyn v co th to IS hai u co th nh v
cung chiu.
- S chuyn v co th to IS hai u co th nh v
ngc chiu.

- Da vo phng thc chuyn v:
+Yu t ct dn: ct khi v tr ny v xen vo v tr
khc.
+Yu t sao chp: sao chp v xen vo v tr mi
(enzym transposase).
+ Yu t sao chp ngc: ARN => ADN v xen vo v
tr mi (enzym transcriptase).
- Cc IS c th vn chuyn
gen bt k khc.
- Transposon co th chuyn v sao chep.
- Chuyn v bao tn.

d. Yu t TnA

- TnA l cc yu t vn ng phc t c cc gen m
ha cho kha nng vn ng ca chng v mang cc
gen chon loc.
- C kch thc ln (khoang 5kb) v phc tp hn cc
yu t Transposon hn hp.

- Cu trc TnA (Tn3: 4957 bp).
+ Ho TnA c on trnh
t lp li hai u 38
40 bp),v tr res gia
v ba gen.
+ Res (resolution site):
v tr ti t hp c hiu.
+ TnpA: transposase.
+ TnpR: resolvase (cht
c ch).

+ ampR (hay bla) m ha
+ -galactamase (xc nh tnh khng vi amp.
- C ch chuyn v ca Tn3
3.2. Yu t di truyn vn ng
eukaryote
-Cc yu t di truyn vn ng eukaryote chia lm 2
nhm l cc retrotransposon v cc DNA transposon.
a. Cc retrotransposon
- Gery Fink v cs. l nhng ngi u tin s dung
nm men nghin cu iu ho hot tnh gene
eukaryote.
- Cc tc gia ny a phn lp c hng ngn t
bin gene HIS4 m ha enzyme tham gia tng hp
histidine.
-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 o chng c c tnh ca retrovirus (virus
ca ng vt).
- C s ging nhau trong cu trc v thnh phn gen
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 goi l trnh t
nm 2 pha on m ha, ca hai u cha gen gag
v gen pol.

- S chuyn v nh retrotransposition

b. ADN transposon
- Nhiu yu t di ng c tm thy eukaryote
chuyn v bng c ch ging vi vi khun.
- Ban thn yu t IS v transposon hoc l ban sao
ca chng c th xen vo v tr mi trn genome.
- Cc yu t di chuyn theo cch ny c goi l yu
t nhm 2 hoc ADN transposon.
- Yu t di ng c McClintok pht hin u tin
ng l cc ADN transposon.
- Tuy nhin tnh c trng phn t ca ADN
transposon u tin li l yu t P rui dm.
a. Yu t P
- P di khoang 2907 bp, cha on lp ngc chiu
hai u di 31 bp.
- P c 4 exon.
- C th chng P c khoang 30 50 yu t P.
Hin tng lon san con lai rui gim:
- P X M =>Con
lai lon san.
- Chng P mang yu
t di truyn khi xm
nhp vo t bo
trng chng M gy
lon sn.
- Trong cc m soma,
hai intron u b ct b
s sn sinh ra protein
c ch (66 kD) hot tnh
ca yu t P.
- Trong cc m t bo
mm, c intron 3 cng
b ct b, sinh ra protein
87 kD, l enzym
transposase.
- S hot ng ca yu t P ph thuc vo cytotype
trng thi yu t ny truyn theo dng m qua t
bo cht ca trng.
b. Yu t Ac v Ds
- ng, yu t Ac v Ds c th di chuyn trn nhim
sc th, tc ng ln cc gen kim tra mu sc ca
cc ht aleuron. Cc yu t ny c phn lp cho
thy c lin quan vi ADN 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 ao ngc m ha cho transposase, yu
t Ds li khng m ha cho transposase.
- Cac ho gen nhay ng:
- C ch gy t gy Ds:
4. Yu t di truyn vn ng ngi








- Ngi c khoang 3000 5000 L1 hon chinh v
khoang 500 000 L1 khng hon chinh (b ct xn u
5. Chi c mt s t L1 hon chinh c kha nng vn
ng.
- Chi co Alu co kha nng chuyn v.
- 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.

- SINE ngi c goi 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 chi
mt phn hoc ton b, chim hn 10% genome ca
ngi.
- Trnh t Alu y c kch thc 200 nucleotide.
ADN genome mang yu t di ng ln hn 20 ln
ADN m ha cho tt ca protein ngi.
VI. TM NG V CU TRC TM NG
- Cu trc centromere hon chinh l 1 phc hp rt
phc tp vi s tham gia ca nhiu loi protein ong
vai tr khc nhau bao gm : ADN li chromosome lin
kt inner Kinetochore v phc hp protein
kinetochore.
- Centromere l mt vng AND thng tm thy gn
gia mt NST, ni 2 chromatid tip xc. N tham gia
vo s phn chia ca t bo, l im gn t v sc.
1/ V tr centromere trn NST:

- Centromere chia th nhim sc thnh 2 vai, chiu
di ca 2 vai phu thuc vo v tr centromere v
phn loi cc th nhim sc.
- T o ta c th phn loi NST theo v tr ca tm
ng thnh: NST tm mt, NST tm lch, NST tm
gia v NST tm cn gia.
Ic=P/(P+Q)
P:chiu di vai ngn
Q:chiu di vai di

A: Th nhim sc tm mt (acrocentric chromosome) c
trung tit u mt ca vai ngn.
B: Th nhim sc cn mt (telocentric chromosome) c
trung tit gn u mt ca vai ngn.
C: Th nhim sc cn tm (tm lch) (submetacentric
chromosome) c trung tit gn chnh gia (vai P ngn
hn vai Q).
D: Th nhim sc tm gia (metacentric chromosome)
c trung tit chnh gia chia 2 vai bng nhau.
a/ Cu trc v trnh t AND-CDE:
- Trong qu trnh nay chi ca nm men
(Saccharomyces cerevisiae), chiu di on AND
centromere (CEN AND) cn thit phn bo l
125bp v c th c chia lm 3 vng trnh t bao
tn CDEI, CDEII, CDEIII.
2/ Cu trc centromere:
- CDE-centromere AND Element- l nhng nhn t c
vng trnh t AND xc nh ca centromere mang
vng trnh t bao tn v trnh t tng ng. C 3
nhn t chnh l CDEI, CDEII v CDEIII.
- Mt nghin cu ca Hieter v cc cng s cho thy s
chon loc chc nng ca CEN AND da vo cc plasmid
ARS mang 1 gen SUP11 tARN.
- Cc plasmid ARS trong nm men S.Cerevisiae c kha
nng t ti to, nhng trong qu trnh phn chia ca t
bo, cc plasmid ARS rt him khi c truyn cho cc
t bo con, do o chng c tch ly t bo m.
- CEN ADN c trong plasmid se gip cho chng c
phn ly ung cch. Nhiu ban sao ca gen SUP11 tARN
se cht v chi c plasmid c CEN ADN sng st.
- V tr trung tm ca tt ca cc CEN ANDs l vng
giu AT, nht l trnh t ca nhn t CDEII (87-98%
A:T v T:A). Vng trnh t ADN giu AT thng c xu
hng thay i v tr sp xp ca trnh t A:T v T:A,
nhng cc nhn t ny khng mang trnh t nhn bit
trnh t bao tn.
- Nh vy, vic thay th 1 phn trnh t CDEII mt
cch ngu nhin trong vng trnh t giu AT lm giam
ang k chc nng ca centromere trong nguyn
phn v giam phn.
- CDEI v CDEIII nm lin k 2 u ca CDEII, l
vng c mc bao tn cao v mang cc trnh t hai
chiu. CDEI gm c vng trnh t c bao tn cao
c chiu di l 8bp c mt ca 16 CEN ADNs
- Trong s tng phan, nhn t CDEIII di 16bp a
cho thy 1 phn tng ng gia 16 CEN AND, tr
vng trung tm c bao tn cao. Phn cn li ca
vng trnh t ny l 1 u giu A:T u 5 v 1 u
giu T:A u 3.
b/ Cu trc cc protein:
- ADN chromosome lin kt inner kinetochore : cha trnh
t bao tn giu trnh t bao tn giu A-T c kch thc
khoang 220bp, 15-20nm. c chia thnh 3 vng.
Vng I II III
Trnh
t
5A(G)TCACA
(G)TG3
>90% A-T 25bp
c bo tn
cao
Chc
nng
Phn li NST v
cc cc TB trong
NP v GP1
ng vai tr
quan trng trong
chc nng
centromere
ng vai tr
quan trng
trong xc nh
cu trc ADN
- Inner kinetochore l phc hp protein trong o c 2
loi c ch nhiu :
+ CENP-A (centromere proteins) : dng bin th protein
H3 thay th cho H3 ca nucleosomes, CENP-A l c
s cho centromere hot ng. Vi t bo thiu CENP-A
s phn chia xay ra khng cn chnh xc.
+ CENP-B l loi protein lin kt vi trnh t lp li
trn ADN ca tm ng. ngi, vng lp li ny
c bit n l alpha satellite ADN.
- Hot ng kinetochore protein t di s kim sot
ca h thng t bo bi nhng phc hp c chc
nng iu ha: APC, phc hp thc y k sau
(anaphase-promoting); CEN, ADN tm ng; SCF,
phc hp ubiquitin-ligase.
+ Ngoi ra cn c s tham gia ca nhiu loi CENP
khc. Ngy nay ngi ta a bit c 20 loi CENP
khc v c th tng hn na trong thi gian ti khi
chng ta hiu r s hnh thnh kinetochore.