t bin gen

Mc tiu : 1. - Trnh by c nguyn nhn v hu qu ca t bin. 2. - Trnh by c cch t bo sa cha ADN h hng trnh t bin. 3. - Nhn din c mt s bnh di truyn ngi do t bin gy ra. 4. - Trnh by c tc ng ca cc yu t gy t bin. 5. - M t c vic s dng cc yu t gy t bin trong phng th nghim v cc phng php chn lc t bin vi sinh vt.

M u
t bin gen: S thay i trong trnh t nucleotid trn gen.

- To ra mt protein t bin vi trnh t acid amin thay i ; - Tc ng n qu trnh iu khin s tng hp ca cc sn phm gen.

Cc loi t bin gen

t bin sinh dng : t bin xy ra cc t bo khng sinh sn - To ra cc thay i c tnh cc b, khng di truyn c (u sc t ngoi da ngi, to ra nt rui); - Nguyn nhn gy ra ung th; - Nguyn nhn ca s lo ha. t bin dng mm : t bin xy ra trong giao t , c truyn cho i sau.

t bin gen (t bin dng mm)

Hu ht l t bin ln (s khng c pht hin nu hp t khng c hai bn ca allel t bin). Pht hin v o tn s t bin cn phi quan st :

- Cc t bin tri ang xy ra trn nhim sc

th thng; - Cc t bin ln v tri trn nhim sc th X.

T l t bin ca cc gen:
C hai gii thch cho s khc nhau v t l t bin ca cc gen:
-

Kch thc ch
im nng

-Kch

thc ch: Gen c vng m ha protein rt ln, c nhiu base c th b thay i hoc lm mt chc nng ca gen.

Th d: - Bnh u x thn kinh (Neurofibromatosis NF): do di truyn gen tri trn nhim sc th thng (l mt bnh di truyn lin quan n ri lon thn kinh nh hng n no, ty sng, dy thn kinh v da. ) Bnh lon dng c Duchenne: t bin trong gen DMD; di truyn ln lin kt vi X; Gen DMD quy nh protein dystrophin protein m bo tnh ton vn ca si c. Nhng b trai vi bnh teo c Duchenne khng sn sinh protein dystrophin trong c bp ca chng. Bnh lon dng c Duchenne chim t l khong 1 trong 3500 ca sinh b trai trn ton th gii.
-

T l t bin ca cc gen:
-im

nng: mt s gen nm trong vng nhim sc th nhy cm hn vi s ph hy/ thay i di truyn, hoc gen c cha cc trnh t d b thay i bi cc t bin t pht hn. V d gen chng lon sn sn c cha im nng (trnh t CpG).

Tn s t bin
Tn s t bin c nh gi trn mt ln sao chp, mt ln phn bo hay trn mt giao t v trn mt t bo trong mt th h. Cc gen khc nhau ca cng mt sinh vt c th c tn s t bin khc nhau nhng tn s t bin t nhin i vi mi gen l n nh. Tn s t bin thng c biu hin: 10-n th d:

E. coli nhy cm vi streptomycin t bin thnh khng

streptomycin vi tn s 4.10-10 = trong 10 t t bo ca mt th h c 4 t bin StrR xut hin ngu nhin.

 ngha ca t bin gen

-

BG c ngha quan trng trong tin ha;

Hu ht cc t bin l ln v nh vy s khng c biu hin trong iu kin d hp t; S thay i cc base c th xy ra trong vng khng m ha ca phn t ADN v cc acid amin b thay i c th khng lm thay i c tnh protein; thay i trnh t nucleotid c th khng biu hin thnh s khc bit tnh trng nhng l cc t bin ln tch t trong b gen.
-

Hu ht cc t bin l bt li

c th gim thiu cc bin i trong chui ADN hoc phc hi trnh t gc bng cch sa cha cc t bin.

ng dng ca t bin gen

-K

thut cm ng t bin: + Trong cy trng v vt nui to ra cc chng mi + Trong cng ngh vi sinh hc hin i to ra cc chng vi sinh vt lm tng nng sut v to ra sn phm mi. - K thut ti t hp ADN cho php chn mt t bin c hiu vo mt gen qui dnh trc (t bin im nh hng) c th to ra cc t bin tt v hu ch.

Cc loi t bin
Cc loi t bin thng gp:

t bin im
t bin lch khung t bin a im

12

t bin im
t bin im : thay i mt base trong chui

ADN do ha cht hoc sai st trong sao chp ADN.

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 delection)

13

t bin im do thay th cp nucleotid

Mt cp nucleotide trong gen c thay th bng mt cp nucleotide khc t bin thay th cp base c chia lm 2 loi: Chuyn v: Ph bin nht S chuyn v, bt cp sai, c th gy bi acid nitr hoc cc ng ng base nh 5-bromo-2deoxyuridin (BrdU). o chuyn

14

Chuyn v
Pyrimidin c thay th bi pyrimidin khc

hoc

CT

purine bi purine khc AG hoc GA

15

Chuyn v Pyrimidin thay th bi pyrimidin khc T C hoc CT

Nguyn thy t bin Ln sao chp tip

AT
GC CG TA

AC
GT TG CA

AT
GC TA CG

+
+ + +

CG
TA GC AT

purine bi purine khc A

Nguyn thy t bin

hoc G
GC
AT CG TA

Ln sao chp tip

AT
GC CG TA

GT
AC CA TG

+
+ + +

TA
CG AT GC
16

t bin li bin

t bin im c khuynh hng c trng l t

bin li (hi bin): ngha l chuyn li dng

nguyn thy. iu ny xy ra v ln t bin tip theo ti cng mt im c mt phn ba c hi quay li chui ban u.

17

o chuyn

pyrimidin c thay th bi purine hay purin bi pyrimidin (C/T A/G).

18

Cc loi t bin im
t bin ln hay cn gi l t bin cng ngha: m ha cho cng acid amin, khng c nh hng n protein cui.
-

t bin lch ngha: m ha cho acid amin khc, c th lm protein khng c chc nng.
-

t bin v ngha: m ha cho codon kt thc, c th lm ct protein.

-

Cc loi t bin im
Le ch ngh a UAC Tyrosine Cu ng ngh a

CAC Histidine

UAG

UAU cu ng Tyrosine

Protein la

Protein kho ng c to ng hp

Protein bnh th ng
20

nh hng ca t bin thay th base

t bin thay th base c th xy ra: trong promoter ; cc vng iu ha 5 ca gen hoc ; trong intron t bin thay th base c th nh hng n s phin m, dch m hoc mc xon bn ca ADN. Nhiu -thalassemia l hu qu ca cc t bin khng-cu-trc ny nh hng mc biu hin cc gen globin. Mc nh hng ty thuc nucleotid b thay th v v tr ca n trong ADN.
21

Ni dung ca t bin cng ngha, lch ngha v v ngha

22

t bin ln (t bin cng ngha)

B m ha cho cng acid amin, khng c nh hng n protein cui. V d codon GCA hoc GCG trong mARN u ngha l

arginin (loi ny thng ng vi t bin chuyn v

v tr th ba - base linh hot ca codon.

23

t bin cng ngha

t bin lch ngha

t bin lch ngha: m ha cho acid amin

khc, c th lm protein khng c chc nng.

V d trong chui protein -globin gy bnh thiu mu hng cu hnh lim: GAG

glutamat

GUG

valin
25

t bin lch ngha

t bin v ngha

t bin v ngha: m ha cho codon kt thc, c

th lm ct protein.
Tc ng ca cc t bin v ngha ty thuc protein b ct bao nhiu v mc cn cho hot ng ca protein. Thng lm mt chc nng protein v gy bnh

27

t bin v ngha

t bin lch khung

Bn cht ca t bin lch khung: do chn hoc mt mt hoc nhiu nucleotid trong vng m ha ca gen. Hu qu: lm thay i khung c Thay i tt c acid amin pha sau t bin v rt d to sn phm khng chc nng v khc hn vi protein bnh thng.

Ngoi ra, cc khung c sai thng cha cc codon kt thc, s lm qu trnh tng hp protein dng sm.
29

Vic thm hoc bt mt nucleotid vo khung c s lm thay i acid amin k t v tr tr i. Nu t bin thm hoc bt cng xy ra trn mt phn t ADN th do trao i cho gia cc base tng ng m t bin m c th sa cha li khung c sai do t bin dng gy ra. Nh ch c acid amin no nm gia hai im t bin b sai m thi. Nh vy, t bin m tc ng nh mt t bin km hm ngoi gen i vi t bin dng hoc ngc li.
30

t bin lch khung c do mt nucleotide

t bin lch khung do thm nucleotide

t bin a im
Bn cht:

Nhng thay i tc ng n hn mt base;

Thay i t hai n hng ngn base, nhng thng nht l ch vi base. Hu qu: thng l xo b mt phn ca trnh t m ha.

t bin a im khng c tnh hi bin, ngha l t bin c n nh.

Phn bit t bin im v t bin a im v nguyn nhn v hu qu

t bin im t bin a im

Nguyn Thay i mt base trong chui Nhng thay i tc ng n

nhn

ADN do :

hn mt base:
base, nhng thng

- s chuyn v, pyrimidin c Thay i t hai n hng thay th bi pyrimidin khc ngn hay purine bi purine khc (A G, C T) - o chuyn pyrimidin c thay th bi Do gen nhy hoc cc li khi sao chp ca cc yu t lp li nht l ch vi base

purine hay purin bi pyrimidin

(C/T A/G)

(v d cc lp li AT)

34

Phn bit t bin im v t bin a im v nguyn nhn v hu qu

t bin im t bin a im

Hu qu t bin cng ngha: Khng c Xo b mt phn ca trnh t m

nh hng n protein cui

ha dn n cc hu qu:

t bin lch ngha: Codon S xa mt on ln s lm mang t bin m ha cho acid thay i hon ton protein to

amin khc , c th lm protein ra,

khng c chc nng xa b on ngn cng gy t bin v ngha: Codon mang hu qu nghim trng nu n t bin m ha cho codon kt lm thay i khung c (nu s thc c th lm ct protein. lng base b xa khng chia ht cho ba).
35

Nguyn nhn t bin

- t bin t nhin (xy ra mt cch t pht) ;

- t bin cm ng gy bi cc tc nhn t
bin.

t bin t nhin

Xy ra trong t nhin mt cch ngu nhin ; Xy ra vi tn s nht nh; Khng xc nh c ngun gc.

C ch Do s gn sai base trong qu trnh sao chp ADN s to ra mt t bin im. T l t bin t pht l 1/107 bn m sao ca gen s c mt t bin im, ngha l 10-7 t bo s c t bin v gen .

Cc dng t bin t nhin:

- h bin, - kh amin, - lch khung, - v oxy ho ph hu do cc gc t do.

38

1. H bin

Bn loi base ca ADN c th tn ti dng h

bin, tc l kh nng hon v gia hai dng keto v

enol; amino v imino.

Nu trong khi ADN sao chp, G dng enol, polymerase s thm T vo thay v bnh thng l C + Nh vy G C chuyn v thnh G T + Khi sao chp mch T, s l A - T Kt qu l s chuyn v G-C thnh A-T gy t bin chuyn v. Dng imino (=NH) him gp, nhng cng gy bt cp nhm.
39

Th d 1: 5-Bu (5-Br Uracyl) c th bt cp vi Adenin to A = 5Bu (dng keto). Nhng khi 5-Bu h bin sang dng Enol (5-Bu*) th 5Bu* li bt cp vi Guanin to G 5Bu*. T mch khun, G c sao chp thnh G C, nh vy A = T chuyn thnh G C.

40

Th d 2: Guanin c th bt cp sai vi 5-Bu dng enol to: G 5Bu*. khi Bu* h bin thnh Bu. T mch khun G sao chp thnh A = Bu T mch khun A sao chp thnh A = T, nh vy: G C thnh A = T.

41

2. Kh amin
Cc phn ng kh amin thng gp l cc phn ng thy phn, lm cho: Cytosin Uracil CH3 Cytosin Thymin Adenin Hypoxanthin S bin i ny dn n bt cp sai. Th d: cytosin bnh thng bt cp vi Guanin to C G. nu cytosin b kh thnh Uracil th c th bt cp vi Adenin. Do : CGU=AA=T
42

C G U =AA=T

43

(a) Thymin (R=CH3) hay Bromouracil (R=Brom) R H O N H

(b) Uracil R H O N H H Adenin N N N N H H

N Deoxyribose O Keto

N Deoxyribose O

N Deoxyribose

Kh amin ho a o ng pha n ho a H R H O N H H R N N H H H O N N N H N H

N Deoxyribose O Enol

N Deoxyribose O Cytosin

N Deoxyribose Guanin
44

3. Lch khung (chn hoc mt nucleotide trn si ADN)

Thng l li ca ADN polymerase khi sao chp cc

on lp li ca mt nucleotide.
Th d: proflavin, cam Acridin, Ethidium bromide,

lm dn si i v ADN polymerase b nh la
chn base vo v tr i din nhiu hn bnh thng.

45

4. Oxy ha ph hy do cc gc oxy Cc gc oxy tng ln trong t bo do chuyn ha oxy ha (hay do phng x).

sn phm oxy ha quan trng l 8-hydroxy

Guanin, gy o chuyn G C thnh T = A.

46

t bin cm ng do cc tc nhn gy B
Cc tc nhn gy t bin:

- lm tng tn s t bin cao hn mc t nhin

-

c th lm thay i cu trc hoc trnh t ADN l tc nhn vt l: phng x, tia X, tia t ngoi, Tc nhn ha hc: cc ng ng ca cc base

nit, acid nitr (HNO2), cc cht alkyl ha mnh.

Tc nhn ha hc gy t bin
Tc nhn gy t bin ha hc c chia lm 4

nhm :

Nhm base ng ng Cc cht ha hc thay i cu trc v c tnh bt cp ca cc base, gm cc cht kh amin v cc cht alkyl ha

Nhm chn vo ADN (tc nhn lm lch khung)

Cc tc nhn thay i cu trc ADN

Bn tc nhn ha hc gy t bin
1. Nhm base ng ng: c cu trc ging purin hay pyrimidin c th gn vo ADN ti v tr ca cc base bnh thng khi sao chp. Th d: BU ng ng ca Thymin Aminopurin ng ng ca Adenin Cc tc nhn ny gy chuyn v nh gy h bin t pht.

49

Nhm base ng ng : bromouracil (5Bu) v aminopurine.

A=T
BromUracil (Bu) thymin (Br thay cho CH3-Bu) nn c th bt cp vi Adenin khi Bu h bin t dng Ceto sang Enol (Bu*) c th bt cp vi Guanin to Bu*- G,

A = Bu

G Bu*

A = T c thay bng G C GC
50

(a) A T Sao chp (Mch khun A) Sao chp (Mch khun BU*) G C (b) G C Sao chp (Mch khun G) Bc t bin Sao chp (Mch khun BU) A T Sao chp (Mch khun A) A BU G BU* Sao chp (Mch khun G) G BU* A BU

Aminopurin ng ng ca Adenin. Aminopurin c th bt cp vi Thymin hoc Cytosin (him khi) gy chuyn v.

2. Cc cht ha hc thay i cu trc v c tnh bt cp ca cc base, gm cc cht kh amin v cc cht alkyl ha 2.1. Tc nhn kh amin: acid nitr to bi s phn gii cc nitrit (cht bo qun) trong thc phm bin :
- Cytosin thnh Uracil; - Methylcytosin thnh Thymin;

- Adenin thnh hypoxanthin kh amin, hypoxanthin

trong ADN bt cp vi C gy chuyn v.
53

C ch kh nhm amin (NH2) ca cc base nitric gy bt cp sai: RNH2 + HNO2 R-OH + N2 + H2O R-OH bt cp sai Th d cht kh amin nh acid nit (HNO2) kh: Cytosin thnh Uracil; hoc

Adenin thnh Hypoxanthin.

gy bt cp sai

.
54

2. 2. Tc nhn alkyl ha :
Nitrosoguanidin (NTG), S) Methyl methane sulfonat (MMS), Ethyl methane sulfonat (EMS), c th gy t bin t nht bng 3 cch :

Thm nhm methyl (-CH3) hay ethyl (-C2H5) vo Guanin to base ng ng ca Adenin dn n s bt cp b sung sai ; Mt purin do Guanin b alkyl ha to l hng trn ADN, khi sao chp c th lm t mch; Lin kt cho gia cc mch ca mt hoc cc phn t ADN khc nhau lm mt nucleotid.

Guanin H
8

O6ethylguanin C2H5 O H
8

N
7

N
7

O
5 4 3 6 1NH

9 4

5 3

6 1NH

N NH2

NH2

O6ethylguanin C2H5 H
8

N
7

O
5 4 3 6 1NH

Thymin O CH3 H N N H

NH
6

Sba t ca p cu a O ethylguanin v i thymin

Tc nhn ha hc gy t bin

3. Nhm chn vo ADN (tc nhn lm lch khung):

cc cht proflavin, cam acridin, ethidium bromide l cc phn t a vng phng tng tc vi cc base ca ADN v chn vo gia chng.

S chn ny lm gin si i ADN v ADN polymerase b nh la chn base vo i din nhiu hn bnh thng, lm lch khung ADN to thnh.

57

Tc nhn ha hc gy t bin

4. Cc tc nhn thay i cu trc ADN Cc tc nhn ny c th l:

Cc phn t ln (knh cng) gn vo base trong ADN v lm chng tr thnh khng m ha. V d N-acetoxy-2-acetylaminofluorene (NAAAF)

Cc tc nhn gy lin kt cho trong v gia cc si. V d cc psoralen c trong mt s rau v dng trong iu tr mt s bnh da.
Cc cht ha hc gy t si ADN. V d cc peroxid. Cc hydrocarbon a vng. V d cc benzypyrene c trong xng.
58

Tc nhn bc x gy t bin

Tc dng gy t bin ca bc x c 2 c im:

Khng c ngng tc dng tc l khng c liu

lng v hi. S lng t bin t l thun vi liu lng phng x, khng ph thuc vo cng v thi gian chiu x.

59

Cc bc x in t
Bc sng ca cc bc x ny thay i ty theo loi

bc x v t l nghch vi nng lng ca chng.

Cc sng di nht (radio AM) c nng lng t nht cc sng ngn hn l FM, TV, viba, hng ngoi, kh kin, tia t ngoi (UV), tia X v tia gamma ln lt c nng lng gia tng.

Tia t ngoi (cn gi tia cc tm) c nng lng

phng x cao nn c ngha sinh hc.
60

Bc x ion ha

Tia X, tia gama, ht phng x v c th lm ion

ha cc phn t trn ng i ca chng v gy bin

i ADN do c tc ng mnh n cc phn t sinh hc.

Bc x UV khng ion ha nhng c th phn ng vi ADN v cc phn t sinh hc khc, v cng l tc

nhn t bin quan trng: Tia t ngoi lm tng mc

nng lng ca nguyn t, thng to dimer pyrimidin (gn hai thymin k nhau ca mt mch.)
61

Ngun bc x
-Bc

x t nhin sn xut bc x nn.

- Ngun bc x t nhin : + cc tia v tr t mt tri v khng gian; + t cc nguyn t hot ng phng x trong t, trn mt t v trong khng kh (radon).

- Mc bc x nn thay i ty vng a l.

Tc dng sinh hc ca bc x
Bc x ion ha sn sinh cc gc t do;

Cc gc t do s tng tc vi ADN, protein, lipid

trong mng t bo, v.v c th gy h hng ADN v protein, c th lm hng c quan, ngn phn chia t bo hoc lm cht t bo.

nh hng di truyn ca bc x

1. Bc x ion ha c th gy:

t mt hoc c hai si (c th dn n sp xp

li, xa mt, mt nhim sc th, t bo cht nu

khng sa cha).

Ph hy cc base (t bin). Lin kt cho trong ADN hoc ADN vi protein.

2. Vai tr gy t bin ca tia UV

Bc x UV c u tin hp ph bi cc base ca ADN v cc acid amin thm ca cc protein, v th n c nh hng sinh hc v nh hng di truyn quan trng. UV c phn loi theo bc sng: - UV-C (180 - 290 nm): st trng mnh nht, gy cht, khng c trong nh sng mt tri v n c hp ph bi tng ozon. - UV-B (290-320 nm): phn on gy cht/t bin chnh ca nh sng mt tri - UV-A (320 nm - kh kin) l cn UV, cng c tc dng xa mt (v n to cc gc oxy) nhng n sn xut rt t dimer pyrimidin.
65

UV-B v UV-C: to cc dimer pyrimidin trong ADN, y l lin kt ng ha tr gia cc pyrimidin k nhau trong mt si
Cc dimer ny lm ngn chn s phin m v sao chp ADN v gy cht nu khng sa cha. Chng c th kch thch t bin cng nh sp xp li nhim sc th.

Di tc ng ca tia t ngoi, cytosin gn thm phn t nc vo lin kt C=C ca mch vng v thymin b t lin kt C=C mch vng, ni 2 phn t thnh thymin dimer.
66

Tc ng ca tia t ngoi ln cc pyrimidin (a) Dimer thymin-cytosin (b) Dimer thymin-thymin

(a) T O CH3 N + O (b) N T O CH3 N + O N N O PR O N N O
67

C NH2 N N T O H3C N UV N O UV PR O N

TC O CH3 N N TT O CH3 CH3 N O N O NH2

Cc c ch chng li t bin
o nghch sai hng: enzym hot ng phc hi cu trc bnh thng khng c gy

khung.
Loi b sai hng: ct b v thay th base hoc phn nucleotid sai hng hoc khng thch hp. Dung np sai hng: khng sa cha tht s, chp sai hng v tip tc sng.
68

o nghch sai hng

3 c ch thng gp: Quang phc hi (photoreactivation) Lm mt nhm alkyl Ni cc ch t si ADN

69

o nghch sai hng = quang phc hi.

Bn cht ca quang phc hi:

enzym phospholyase xc tc phn ng ct cc

dimer pyrimidin c to ra di tc ng ca tia UV bng cch lm t cc lin kt ng ha tr. Hin tng quang phc hi l mt c trng trong tc ng ca tia UV. Sau khi chiu tia t ngoi ln t bo, nu ngoi nh sng, th cc sai hng phn ln c phc hi. nh sng c tc ng hot ha enzym phospholyase, ct t cc thymin dimer.
70

T DNA UV

PR T DNA vi dimer T

Enzym photolyase xc tc phn ng ny, c nhiu trong

vi khun, t bo nhn tht bc thp, cn trng v thc

vt, khng c trong ng vt. Gen phr c ng vt c v nhng m ha cho protein c chc nng h tr loi sa cha khc.
71

Sa sai bng cch lm mt nhm alkyl (dealkylation)

Enzym O6-methylguanin-ADN methyltransferase chuyn nhm alkyl t nguyn t oxy trn ADN sang gc Cystein trn enzym ny lm enzym ny mt hot tnh. Phn ng khng thun nghch. Cc li c sa cha gm O6-methylguanin, O4methylthymin, v cc methylphosphotriester gy ra bi cc tc nhn alkyl ha nh N-methyl-N-nitroN-nitrosoguanidine (MNNG). Enzym ny cng sa cha cc guanin b alkyl ha khc nh O6-ethylguanin v O6-butyl-guanin, mc d hiu sut thp hn nhiu. Cc MTase c tm thy tt c sinh vt c nghin cu.
72

Th d E. coli:
C hot tnh
Cys SH

Bt hot
Cys S CH3

OCH3 N H2N
6

O N N R N N N R

Methyltransferase
H2N

O -Methylguanin

Guanin

Enzym O6-methylguanin-ADN methyltranferase chuyn nhm alkyl t nguyn t oxi trn ADN sang gc Cystein trn enzym.

73

Sa sai = Ni cc ch t ca si n

ADN ligase c th nhanh chng ni li cc si ADN b t do tia X hay ha cht nh peroxyd gy ra.

Chng li B = Loi b sai hng

Theo cc c ch:

Ct b base sai
Sa cha lch i

Sa cha bng ct nucleotid NER

(nucleotide excision repair)

75

Loi b sai hng = Ct b base sai

Enzym N-glycosylase nhn bit cc base sai v thy gii lin kt N-glycosilic (ni base vi ng pentose) to ra v tr khng c purin hay pyrimidin (AP).

Sau , endonuclease ct b AP v ADNpolymerase chp y ch trng da vo khun mch b sung i din v c ligase ni li.

76

N-glycosylase nhn bit cc base sai v thy gii lin kt N-glycosilic to ra v tr khng c purin hay pyrimidin (AP).

Endonuclease ct b AP

ADN-polymerase chp y ch trng da vo khun mch b sung i din v c ligase ni li.

77

Loi b sai hng = Sa cha lch i

Mt nhm protein c th qut ADN va mi c

sao chp tm cc base bt cp khng chnh xc

(hoc base khng bt cp) lm sai lch hai chiu xon kp. Base khng chnh xc s c loi b v ADN polymerase s hot ng ln th hai to trnh t ng.

78

Sa cha bng h thng NER (Nucleotid Exision Repair)

H thng NER gm:

Endonuclease ct si ADN cha sai hng c hai

pha.

Exonuclease loi b on ADN ngn

ADN polymerase lp nucleotid vo khong trng

79

DNA vi dimer

Dimer c nhn din v ADN b ct

Dimer b loi b

Khong trng c lp nh ADN polymerase

Khe h c hn li nh ADN ligase

80

81

Chng li B = Dung np sai hng

Sa cha bng ti t hp
Sa cha bng t bin

82

Sa cha bng ti t hp
Ti t hp tng ng: y l c ch sa

cha lp khuyt si con khng dimer da vo

s tng ng ca si nhim sc th em trong qu trnh trao i cho.

Ti t hp khng tng ng: y l phn ng lin kt u khng tng ng sa cha u

ADN b t, gy do bc x ion ha hay cht gy

t bin.
83

Sa cha sai hng bng ti t hp

Sa cha t bin bng ti t hp

85

Sa cha bng t bin

ADN polymerase b ngn ti dimer lm thay i

tnh c hiu ca enzym, nh n c th chn

bt c nucleotid i din no vo v tip tc sao chp.

86

Sa cha sai hng bng t bin

87

Sa sai nh h thng SOS

H thng sa cha ny c kch hot nu cc c

ch trn b qu ti bi cc t bin lan rng.

Trong trng hp ADN b tn thng ngng sao chp, th phn ng SOS hi phc sao chp v chuyn sai hng thnh sa sai trong khi nhn i.

88

Cc tnh trng t bin v protein t bin

Bn cht ca t bin gy hi chng Alkapton niu

t bin gen m ha cho enzym homogentisate 1,2-dioxygenase (HDG) = homogentisate oxidase. t bin hay gp nht l thay valin cho methionin v tr 368. S t bin ny gy thiu enzym homogentisate oxydase trong gan nn khng phn hy c acid homogentisic (tn c l alcapton) thnh mt sn phm khng mu l acid acetoacetic.

Acid homogentisic tch t li trong nc tiu v c mu en khi gp khng kh.

Bnh do gen ln, nm trn NST thng t bin
90

Chui phn ng sinh ha ca bnh Alkapton niu v Phenylketon niu, hai s ri lon di truyn ca s thoi qu acid amin thm

91

t bin sinh dng vi sinh vt

Th VSV pht trin c trn mi trng ti thiu (c cha nguyn ng, ion NH4+ v mt vi ion cn thit khc). dng Tc Chng thu c khng pht trin c trn mi trng nhn gin. gy t bin Th khuyt dng Phi thm vo mt cht khc, thng l acid amin, vitamin nucleotid hoc mt phn t vi lng no .

- Th nguyn dng c th tng hp tt c cc thnh phn t bo - VSV khuyt dng c th do mt hot tnh enzym tng hp cc phn t phc hp.

t bin v bnh ngi

t bin gen v ung th c ch phn t, ung th lin quan n cc bin i di truyn trn ADN lm tn thng tin trnh tng sinh bnh thng. Cc nhm gen lin quan ti ung th - Th nht, cc bin i lm tng hot gen kch thch, kiu t bin ny c hiu qu tri. Allel bin i c gi l oncogen (gen ung th), cn allel bnh thng l proto-oncogen (gen tin ung th). Th hai, cc bin i lm bt hot gen km hm, kiu t bin ny thng c hiu qu ln. Gen km hm c gi l gen c ch khi u.

Gen ung th v gen tin ung th

Cc gen ung th do retrovirus ti np
-Cc

retrovirus bng con ung sao chp ngc tch hp c ADN ca mnh vo nhim sc th t bo ch. -Chng c th ngu nhin mang cc gen ung th lm bin i t bo ch bng cch xen on ADN ca mnh cnh gen tin ung th ca t bo ch. - Cc gen tin ung th hu nh lin quan n cc h thng tn hiu ca t bo nh cc protein, cc th th xuyn mng, cc gen iu ho, .

Cc cch bin i gen tin ung th thnh gen ung th

Ba cch ch yu :

- Mt on hoc t bin im trong trnh t m ha

- Khuch i gen - Cu trc nhim sc th Cc gen c th b bin i do t bin im, mt on, chuyn on nhim sc th hay do xen on

b gen ca retrovirus

Ba cch bin i gen tin ung th thnh gen ung th

Ma t oa n hay o t bie n ie m trnh tma hoa

Khue ch a i gen

Ca u tru c la i nhie m sa c the

ADN ARN

hay

Protein ta ng hoa t c ta o ra v i so l ng bnh th ng

Protein bnh th ng c sie u sa n xua t

Ca c enhancer ma nh ke ca n la m protein bnh th ng c ta ng sa n xua t

Sdung h p v i gen phie n ma ma nh la m protein dung h p c ta ng sa n xua t hoa c protein dung h p ta ng hoa t

25. Cc h thng chn lc t bin vi sinh vt

Cc h thng chn lc t bin gip pht hin cc t bin him hoi trong ngun t bin rng ln. Cc h thng chn lc t bin vi sinh vt hon thin cc phng php chn lc t bin trong phng th nghim. Phng php khng: Dng thuc hoc phage chn lc vi khun.

97

t bin khuyt dng

Phng php lm giu chm:

- Dung dch vi khun pha long c cy trn b mt mi trng thch ti thiu mc thnh cc khun lc ri. - Ph mt lp mng mi trng tng t ln mt mi trng thch va ri, cc khun lc bnh thng mc ln.
- Sau li ph thm mt lp mi trng dinh dng c b sung v tip cho cht b sung khuch tn. - Cc t bin khuyt dng s mc sau khi c cht b sung, nn khun lc nh hn do mc chm.
98

Phng php lm giu hn ch

Cc vi khun c cy trn mi trng ti thiu c

mt t cht b sung. Trong iu kin ny, cc t

bin khuyt dng mc n khi ht cht b sung th dng, do to khun lc nh.

Cc vi khun bnh thng tip tc to khun lc to.

99

Phng php lm giu nh penicillin

Penicillin c tc ng dit khun dng phn chia. Vi khun c cho vo mi trng ti thiu c penicillin. Cc vi khun ang tng trng b dit, ch cc t bo t bin khng tng trng cn sng st.

Sau , hn hp c cy ln mi trng khng c

penicllin th cc t bin mc ln vi t l tng i cao hn.

100

Phng php lc
Dng chn la cc t bin khuyt dng nm si. Dung dch bo t c nui trong mi trng dinh dng thiu cht b sung. Cc t bin thiu cht b sung khng mc c, cc dng bnh thng mc ra thnh nhiu si. Khi lc qua mng lc si thy tinh, cc dng bnh thng nhiu si b gi li, cc dng t bin i qua mng lc. Dung dch c nhiu dng t bin c cy trn mi trng c cht b sung v kim tra tm cc dng t bin.

101

Phng php in
a go c tre n mo i tr ng a y u
(a)

(a) a gc trn mi trng y . (b) Khc g bc ming nhung. (c) Mi trng c b sung cht A v B. (d) Mi trng c b sung cht A, B v C. (e) Mi trng c b sung cht A v C
102

Khu c go
(b)

Mie ng nhung

Ca c ba n sao tre n mo i tr ng co bo sung

(c) (d) (e)