C ch phn t ca qu trnh ti bn DNA

Nhng nghin cu tip theo tm ra c ch phn t ca qu trnh ti bn. l mt qu

trnh phc tp c s khc bit prokaryote v eukaryote nhng phi tri qua c ch chung
sau:

Cc lin kt hydrogen n nh cu trc xon v lin kt hai mch n vi nhau phi

c ph v tch ri hai mch.

Phi c on mi (primer), tc l on DNA/RNA ngn, bt cp b sung vi mch khun

to u 3OH t do.
Nguyn liu tng hp DNA l cc desoxynucleosid 5-triphosphate (dNTPs): dATP, dGTP,

dCTP v TTP.
Mch khun lun c c theo chiu 3-5 trong khi mch mi c tng hp theo

chiu 5-3. Mi nucleotide mi c gn vo u 3OH ca mch ang ko di bng lin kt

cng ho tr. Nng lng cho s polymer ho n t vic thy phn cc dNTPs, loi ra cc
pyrophosphate.
Mi bc c thc hin mt cch nhanh chng, chnh xc di s iu khin ca
enzyme c hiu.

n v ti bn, im khi u v im kt thc ti bn

Mi on DNA c ti bn nh l mt n v ring l c gi l mt n v ti bn
(replicon). Cc DNA dng vng, kch thc nh (chng hn nh nhim sc th ca vi khun
v DNA virus) ch gm mt n v ti bn duy nht. S ti bn khi u t mt im gi l
im khi u sao chp (origin) v lan ra theo hai hng, hnh thnh hai chc ba ti bn
(replication fork) cho n khi gp nhau ti im kt thc (terminus). im khi u ti bn c
khuynh hng giu A-T d dng khi u tch ri hai mch n.

 eukaryote, mi phn t DNA bao gm nhiu n v ti bn. Mi n v ti bn c im

khi u ring v s ti bn cng lan ra theo hai hng. Khi cc chc ba ti bn gp nhau
th s ti bn DNA c hon thnh. V d nh t bo hu nh in hnh c t 50.000
100.000 n v ti bn, mi n v ti bn c kch thc khong 40 200 kb.
1.4 S tng hp mch mi ca DNA xy ra lin tc trn mt mch v gin ontrn mch
kia(Semi-discontinuous replication - Ti bn bn gin on)

Trong ti bn bn bo tn, c hai mch mi ca DNA c tng hp ng thi mi chc

ba ti bn. Tuy nhin, c ch ti bn DNA ch cho php s tng hp theo hng 5 3, v
th trn hai mch khun c hng ngc nhau, s tng hp mch mi khng din tin
ging nhau.

T im khi u ti bn, trn mch khun 3- 5 s tng hp mch mi din ra mt

cch lin tc theo chiu 5- 3 cng chiu vi hng tho xon. Mch ny c gi l mch ti
(leading strand).
Trong khi trn mch khun 5 3, s tng hp mch mi cng din ra theo hng 5
3 nhng ngc vi hng tho xon. Chnh v th, s tng hp mch mi khng xy ra lin

tc m di dng nhng on ngn c gi l on Okazaki (kch thc 1000 2000 bp

prokaryote v 100 200 bp eukaryote). Mch mi c tng hp theo kiu gin on ny
c gi l mch chm (lagging strand).

Trn thc t, s tng hp c hai mch theo cng hng v mch khun chm c un
vng quay 180o ti chc ba ti bn, tr nn cng hng vi mch khun ti.

Mi RNA
Mch ti v tt c nhng on Okazaki ca mch chm ch c th c tng hp bng
cch ko di mt mi bt cp sn trn mch khun. Mi l mt on RNA ngn, c
tng hp bi mt phc hp protein gi l primosome bao gm nhiu protein v mt enzyme
c tn l primase. V d, E. coli, phc hp primosome bao gm cc Dna B helicase (tch
ri hai mch n) v DNA primase (tng hp mi RNA t khun DNA).
Cc mi RNA ny sau s b phn hy bi RNase v c thay bng trnh t DNA nh
vo hot ng ca DNA polymerase. Enzyme ligase s ni lin cc on DNA li vi nhau.
C ch dng mi l RNA thay v DNA dng nh lin quan n tnh chnh xc cao trong ti
bn DNA v hot tnh sa sai ca cc DNA polymerase.

Ti bn DNA prokaryote
Cc nghin cu v ti bn DNA c tin hnh da trn cc h thng th nghim in vitro.
theo di ti bn DNA, ngi ta dng thymidine H3phng x. Qu trnh ti bn xut pht t
im khi u v lan ra hai pha. Khi quan st DNA vng trn ang ti bn, ta thy dng
DNA hnh con mt (eye replication). Hai chc ba ti bn lan dn, cui cng to thnh hai
phn t DNA lai, trong c mt mch mang du phng x T-H 3. C trng hp, s ti bn
ch xy ra v mt pha.

Din bin ti bn nhim sc th vng trn c th chia lm ba giai on: khi u

(initiation), ko di (elongation), kt thc v phn chia (termination & segregation).

Giai on khi u
Tho xon phn t DNA
M hnh phn t ca DNA vng vi khun hay virus cho ta mt khi nim chung v
cc cu hnh c th c ca mt phn t DNA. Dng th nht l siu xon. Dng th
hai c cu trc li hn, thng l do b t mt ch trn mt trong hai mch ca phn
t DNA. Dng th ba tng ng vi cu trc thng do s ct t trn c hai mch
trong phn t DNA. Trong , dng siu xon l dng c bn v cu trc v chc
nng.

Nh vy, bt u ti bn, phn t DNA phi c tho xon nh vo hot ng ca

cc enzyme c tn l topoisomerase. C hai loi topoisomerase. Loi I tho dng siu
xon, chng gn vo phn t DNA v ct mt trong hai mch. Sau khi gii phng mt
phn t DNA c tho xon, cc enzyme ny s ni li ch t. Enzyme loi I
c bit r nht l protein ca E. coli. Cc topoisomerase loi II c kh nng tho
cc nt ny sinh do cc bin i cu trc ca chui xon kp bng cch ct t c hai
mch DNA. Enzyme c bit r nht l gyrase ca E. coli. Gyrase s dng nng
lng t s thy phn ATP tho xon DNA.

Tch ri hai mch n ti im khi u ti bn

 E.coli, OriC cha bn v tr gn vi protein khi u c tn l Dna A. Mi v tr c

kch thc 9bp. S tng hp cc protein ny gn lin vi tc tng trng t bo v
th vic khi u ti bn DNA cng gn lin vi tc tng trng. tc tng
trng cao, cc nhim sc th ca vi khun c th bt u ln ti bn th hai trc
khi ln ti bn th nht kt thc ti hai im khi u mi. V vy, mi t bo con s
nhn c mt nhim sc th ang c ti bn mt phn.
Qu trnh ti bn bt u khi DNA OriC qun quanh mt phc hp protein Dna A gm
30-40 phn t, mi phn t gn vi mt ATP. iu ny thc y s tch ri hai
mch ti ba trnh t lp 13bp giu A T, cho php protein Dna B gn vo. Dna B l
mt DNA hehicase, nm trong phc hp primosome. Cc helicase ph v lin kt
hydro gia cc base nh nng lng thy phn ATP. C nhiu loi helicase cng hot
ng ng thi: Mt s gn trn mch 3 5 nh cc Rep, s khc gn trn mch 5
3 nh helicase II v III.
Cc mch tch ri s c n nh di dng mch n nh cc protein SSB
(Single Strand Binding lin kt vi mch n). Cc protein ny gn ln khp phn
mch n lm cho hai mch khng kt hp tr li c. Mch khun c s dng
n u th cc protein SSB c gii phng khi khun n .

Giai on ko di
Tng hp mi RNA

Cc DNA polymerase ch c th tng hp DNA bng cch ko di mt mi RNA bt

cp sn trn khun. Nh bit, mi ny c tng hp nh hot tnh ca enzyme
primase c trong phc hp primosome.

S tng hp mch mi din ra theo kiu bn gin

on, ko di mi RNA

DNA polymerase III l mt phc hp dimer. Mi phn gm nhiu n v gn vi nhau,

chu trch nhim tng hp mt mch n DNA nhm m bo cho tc tng hp
ca c hai mch bng nhau. n v c hot tnh polymerase tht s. n v c
chc nng c sa nh hot tnh exonuclease 3- 5, t lm tng tnh chnh xc
trong ti bn. n v gip gn polymerase vo DNA. y l nhn t duy tr, khc
nhau hai mch khun, quy nh di ca on DNA c tng hp mch ti
(di) khc vi mch chm (ngn).

DNA polymerase III bt u ti bn trn mi mch khun bng cch gn vo mch

(nh n v ) v lp cc nucleotide b sung vo v tr tng ng, ko di on mi
RNA bt cp sn trn khun t u 3OH t do ca mi (nh n v ). Cc DNA
polymerase c tnh c hiu cao, ch thm nucleotide vo u 3OH ca mch ang
tng hp.
Ngoi chc nng polymer ha theo hng 5 - 3, DNA polymerase III cn c kh nng
sa sai nh hot tnh exonuclease theo hng 3- 5. Exonuclease l hot tnh enzyme
ct DNA t u mt mt mch. Trn ng di chuyn tng hp mch mi, nu gp
ch nucleotide va lp sai v tr, DNA polymerase III s dng hot tnh exonuclease 35 ct li li b nucleotide sai v lp ci ng vo ri tip tc ti bn (n v ).
Qu trnh ti bn DNA E.coli din ra vi tc nhanh, c th n 50.000
nucleotide/pht.

Hon chnh si mi tng hp

Trn mch chm, sau khi mi on DNA c ko di, mi RNA b di i nh hot
tnh exonuclease 53 ca DNA polymerase I v s b enzyme RNase H phn hy. Cc
ch trng m mi li s c thay bng trnh t DNA mt cch chnh xc nh kt
hp hot tnh polymerase 5 3 (ko di u 3 ca on Okazaki trc) vi hot tnh
exonuclease 3 5 (c v sa sai) ca DNA polymerase I. Cui cng, ligase s ni
tt c cc on DNA trn mch mi tng hp li vi nhau.

Giai on kt thc ti bn v phn chia t bo

Hai chc ba ti bn s gp nhau khong 180 o i din vi OricC. Quanh vng kt
thc ny c vi im lm dng li s ti bn bng cch gn vi mt sn phm ca
gen tus, l mt nhn t km hm hot ng helicase ca Dna B.
Khi s ti bn hon tt, hai phn t DNA vng vn cn dnh vi nhau. Mt
topoisomerase loi II c tn l topoisomerase IV s tch ri chng sau hai
nhim sc th con s c phn phi vo hai t bo con.

Ti bn DNA eukaryote
S ti bn eukaryote vn cn nhng iu cha tng tn nhng cc d liu thu c cho
thy h thng ny kh gn vi h thng ti bn prokaryote. Khc bit ch yu l cc loi
DNA polymerase tham gia vo qu trnh.

im khi u ti bn v giai on khi u:

V s phc tp trong cu trc ca cht nhim sc, tc di chuyn ca cc chc ba ti bn
eukaryote ch t khong 50 bp/giy. Vi tc ny, nu ch s dng hai chc ba ti bn,
phi mt 30 ngy mi ti bn xong mt phn t DNA nhim sc th in hnh ca ng vt
hu nh vi kch thc 105 kb. Do cn phi c nhiu n v ti bn trong mt t bo, in
hnh nh ng vt hu nh l 50.000 100.000 n v ti bn.
eukaryote, c khong 20 50 n v ti bn khi u cng lc ti mi thi im, xuyn
sut phase S. Phn ti bn sm nht ch yu bao gm nguyn nhim sc cht
(euchromatin), tc l phn DNA c hot ng phin m. Trong khi , d nhim sc cht
(heterochromatin) c hot ha mun hn v phn DNA ca tm ng (centromere) v
on cui nhim sc th (telomere) c ti bn sau cng. iu ny phn nh kh nng
p ng khc nhau ca nhng cu trc nhim sc cht vi nhn t khi u.
Cc trnh t khi u ng vt hu nh cha c phn lp nhng ngi ta tin rng s
khi u ca mi n v ti bn c l din ra ty bn trong mt vng khi u c th di
n vi kb v l mt phn ca trnh t DNA lp li trung bnh.
Khc vi prokaryote, cc n v ti bn ca eukaryote ch khi u mt ln trong mi chu
trnh t bo. Mt protein gi l nhn t cho php (licensing factor) cn thit cho s khi u
v ti hot ha ch c kh nng tip cn vo nhn khi mng nhn tan i, v th ngn chn
c vic ti khi u trc hn nh.

Chc ba ti bn v h thng cc enzim ti bn

Trc khi ti bn, DNA phi c tho khi nucleosome ti mi chc ba ti bn. iu ny
lm chm s di chuyn ca chc ba, ch cn 50 bp/giy. Sau khi chc ba ti bn i qua,
cc nucleosome c ti lp rp t cc histone c v histone mi c tng hp. Kch
thc nh ca cc on Okazaki (v d 135 bp SV40) phn nh lng DNA c tho
khi mi nucleosome khi chc ba ti bn i qua. Ngoi s nhn i DNA, cc histone cng
phi c nhn ln gp i trong sut phase S.

H thng cc DNA polymerase eukaryote phc tp hn so vi prokaryote, bao gm:

Polymerase /primase: C chc nng tng hp mi cho mch ti v cho c nhng on

Okazaki ca mch chm. Polymerase ny tip tc ko di DNA nhng nhanh chng b thay bi
polymerase trn mch ti v polymerase trn mch chm. Polymerase khng c hot tnh
exenuclease.
Polymerase : C chc nng ging DNA polymerase I prokaryote, ngha l tng hp i
km vi sa sai v hon chnh mch mi sau khi mi RNA c loi b.

Polymerase v polymerase : C chc nng ko di DNA. Trong kh nng tng

hp on DNA di nht thuc v polymerase vi s tr gip ca PCNA. C hai enzyme ny
u c kh nng c v sa sai.
: c tm thy trong ti th, chc nng cha r.Polymerase

Ngoi cc polymerase k trn, h thng ti bn eukaryote cn c s tham gia ca nhiu

protein chuyn bit nh PCNA (Proliferating Cell Nuclear Antigen - khng nguyn trong t
bo ang phn chia) c chc nng hot ha cc polymerase v , cc nhn t ti bn A v
C (Replication Factor, RF -A, RF - C) cn cho hot ng ca cc polymerase v ...

Ti bn on cui nhim sc th (telomere replication):

Hai on cui ca nhim sc th dng thng khng th c ti bn y bi v khng th
ko di DNA t u 5 ca mch mi tng hp sau khi loi b mi RNA. Do thng tin
di truyn s mt dn khi DNA sau mi t ti bn.
gii quyt vn ny, on cui mi nhim sc th eukaryote bao gm hng trm bn
sao mt trnh t n gin, khng m ha (v d nh TTAGGG ngi) vi u 3 nh ra so
vi u 5.
Telomerase l mt enzyme duy tr di ca telomere. Enzyme ny c cha mt phn t
RNA ngn. Mt phn trnh t ca RNA ny b sung vi trnh t lp ca telomere, do n
c th hot ng nh mt mch khun ko di cc trnh t lp t u 3 t do ca
telomere bng cch tng hp ri dch chuyn lin tip nhiu ln.
Cc eukaryote n bo, chng hn nh nm men, nh vo hot ng ca telomerase m
duy tr telomere, t bo v t bo khi s mt thng tin di truyn. Th nhng cc sinh
vt eukaryote a bo, cc t bo sinh dng, gen m ha telomerase b km hm. V th,
qua mi t phn bo, cc nhim sc th ngn dn i cho n khi chm ti cc trnh t m
ha ca DNA, t bo s gi i v cht. Trong nui cy t bo sinh dng, hu ht cc t bo
ch phn chia 30-40 hoc 50 ln ri cht.
cc t bo ung th, s ti hot ha hot ng ca telomerase khin cho cc t bo ny
c kh nng bt t.

Mt trong nhng tnh cht cn bn ca DNA l kh nng t sao chp (replication) hay l t
nhn i (self duplication). Ngha l t mt phn t DNA m sao chp to ra hai phn t
DNA con ging y nh phn t DNA m ban u.
S ti bn DNA l mt trong nhng tnh cht quan trng, nh m thng tin di truyn c
truyn t th h ny sang th h khc.

L thuyt v sao m DNA theo khun

Gi thuyt ca Watson - Crick

Sau khi xy dng m hnh cu trc ca phn t DNA, Watson v Crick thy rng, nu hai
si DNA tch ri ra th mt si s lm khun tng hp mt si mi v do cc baz nit
lun c xu hng bt cp vi nhau (A vi T v G vi C). Theo d on ca Watson v
Crick, qu trnh tng hp DNA c th tm tt nh sau
- u tin lin kt hydr s b t ra to hai mch n lm khun.
- Cc baz nit mi s bt cp b sung vi cc baz nit trn si lm khun theo qui tc A
i din vi T v G i din vi C.
- Kt thc qu trnh sao m, t mt si DNA xon kp m s to hai si xon kp con. Mi
si xon kp con c mt si mi tng hp v mt si s lm khun.
Ngay sau khi m hnh c nu ra, nhiu th nghim chng minh c tin hnh xc
nhn d on.

Th nghim chng minh ca Meselson - Stahl (1958)

Th nghim ny nhm mc ch chng minh kiu tng hp theo khun ca DNA nh d
on ca Watson v Crick. Th nghim c thc hin trnE. Coli. Vi khun c nui cy
qua nhiu th h trn mi trng c cha nit ng v nng N15 v nit thng N14 .
Nguyn tc ca th nghim l da trn kh nng phn bit t trng ca hai ng v N15 v
N14 c mt trong cc loi DNA khi li tm trn thang nng CsCl2 . Qu trnh th nghim
c tin hnh nh sau
- Th h u (gi l th h 0): vi khun c nui trn mi trng cha N15 nh vy th
h ny, cc nguyn t nit trong DNA ca vi khun s l N15 . Sau khi nui tin hnh tch
DNA v em li tm trn thang nng CsCl2, nhn thy
+ th h 0: ch cha 1 gii n c t trng cao. Nh vy tt c DNA ca th h cha m u
cha 2 si vi nit nng.
- Th h 1: Chuyn vi khun th h 0 sang nui trn mi trng cha N14. Thi gian v
iu kin nui tung t nh th h 0. Sau khi tch DNA v em li tm nh th h 0, th
thy
+ th h 1 ch cha 1 gii n c t trng trung bnh. Nh vy mi phn t con th h 1
c 1 si cha nit nng ca b m v 1 si b sung mi tng hp c cu to t nit nh
N14, gi l si DNA lai.
- Th h 2: Tip tc chuyn vi khun ca th h 1 sang th h 2 v cng nui trn mi
trng cha N14. Qu trnh th nghim tin hnh tng t nh cc th h trn. Sau khi li
tm, quan st thy c mt na s lng phn t DNA l phn t lai ging th h 1 v mt
na s lng cn li l phn t DNA vi c hai si u mang N14

M hnh
th nghim ca Meselson-Stah

Th nghim trn l mt bng chng chng minh gi thuyt ca Watson v Crick l ng.
Qu trnh tng hp DNA phi thc hin theo khun, hai mch ban u c tch ra, mi ci
lm khun tng hp mch mi. Kt qu l, mi phn t con u mang mt mch c v
mt mch mi. Kiu sao m nh vy gi l sao m bn bo tn (semi-conservative).

Th nghim chng minh ca Arthur v Kornberg

Cng trong nm ny (1958), Arthur v Kornberg ti trng i hc Washington tinh sch
mt loi enzyme t E. Coli, gi l DNA-polymerase. Enzyme ny xc tc tng hp mt si
polynucleotide t cc nucleotide-3- phosphat.
Nguyn liu dng cho th nghim gm
- Bn loi nucleotide dng triphosphat: dATP, dTTP, dGTP, dCTP.
- Enzyme DNA-polymerase chit t vi khun E.Coli.
- Ion Mg+2 l cofactor ca enzyme, to iu kin thun li cho hot ng ca enzyme.
Tin trnh th nghim c th m t nh sau
Th nghim th nht c tin hnh vi y cc nguyn liu nh gii thiu trn,
trong cc iu kin thun li cho s tng hp si DNA mi, nhng khng c si lm khun.
Th nghim th hai cng c tin hnh tng t nh th nghim 1 nhng c mt si DNA
lm khun. Si lm khun c thu nhn bng cch lm bin tnh v tch mch, sau
a vo th nghim.
H thy rng, th nghim th nht, enzyme DNA-polymerase khng th tng hp si DNA
t cc nguyn liu khi khng c mt ca si khun, vic tng hp si DNA mi ch xy ra

th nghim th hai, khi c mt DNA khun. iu khng nh rng gi thuyt ca Watson

v Crick l ng. DNA c tng hp ch khi c mt si lm khun.

Qu trnh sao m
Cc enzyme v protein c hiu tham gia sao m
y l mt qu trnh phc tp, thc hin sao m cn thit phi c mt ca cc enzyme,
cc protein c hiu v cc iu kin cn thit sau y:
- Lin kt hydro gia 2 mch b sung ca si DNA m phi b ph v v tch ri tng bc
lm 2 mch lm khun.
- Phi c on mi (primer), tc on RNA mch n ngn bt cp vi mch n DNA
khun t v tr bt u ca mi on sao chp.
- C 4 loi nucleotide dng triphosphat (dATP, dTTP, dGTP, dCTP) bt cp b sung
vi cc nucleotide mch khun.
- Mch mi c tng hp theo hng 5P 3OH.
- Mi bc c iu khin bi enzyme c hiu v c thc hin mt cch nhanh chng,
chnh xc.
- C mt ca ion Mg+2 lm cofactor ca enzyme.
- Trong qu trnh sao chp, ngoi cc enzyme c hiu cn c cc protein c hiu.
Cc enzyme v protein c hiu tham gia sao chp gm c
- Enzyme topoisomerase, lm nhim v tho xon v hai pha trn si DNA kp
- Enzyme helicase, s dng nng lng ATP lm t lin kt hydr gia cc baz nit
ca hai mch khun.
- DNA-polymerase I, -II, -III lm nhim v tng hp si mi v sa sai.
- RNA-polymerase, lm nhim v tng hp on mi.
- Enzyme primase, lm nhim v gn mi.
- Enzyme ligase, lm nhim v ni hai nucleotide ng cnh nhau bng lin kt
phosphodiester to mch polynucleotide.
- Enzyme ribonuclease (RNase) lm nhim v ct b mi sau khi tng hp xong mch mi.

Sao chp theo quy lut b sung

Cc protein tham gia sao m gm c

- Protein B, nhn bit im khi u (origin) sao chp trn si DNA kp.
- Protein SSB (Single Strand Buiding), lm nhim v gi hai mch ca si DNA khng cho
chp vo nhau, to iu kin thun li cho qu trnh sao chp.

Qu trnh sao m
Nhn bit im khi s v tho xon DNA
Qu trnh sao m c nghin cu kh k t bo vi khun E.Coli. S sao m bt u khi
protein B c hiu nhn bit im khi s sao m (replication origine) v gn vo trnh t
c hiu . Tip theo, enzyme topoisomerase thc hin tho xon phn t DNA t im
khi s.
Enzyme helicase s dng nng lng ATP ct t cc lin kt hydro gia cc baz bt cp
ca hai mch phn t, tch hai mch to thnh chc ba hnh ch Y, gi l chc ba sao
m (replication fork). C nhiu loi enzyme helicase: c loi gn trn mch, di chuyn v ct
lin kt hydro theo chiu t u 3' n u 5'; c loi gn ln mch, di chuyn v ct lin kt
hydro theo chiu 5' 3'. Sau khi tch ri, hai mch n s c protein lm cng mch
SSB gi khng cho chp li, lm cho trng thi m xon c bn vng. Mi phn t
protein SSB bm vo 8 nucleotide trn mch n, mi chc ba sao m c khong 250 phn
t protein SSB hot ng. Mch khun c s dng n u, cc phn t protein SSB s
c gii phng n .

Tng hp mi

Enzyme DNA-polymerase ch c kh nng tng hp si DNA mi bng cch ni di u

3'OH t do ca mt on mi bt cp sn trn khun theo chiu 3'OH n u 5'P.
Mi l mt on RNA nh chng 10 nucleotide, c tng hp t khun ca si DNA. Phc
hp enzyme primase- RNA-polymerase bm vo mch n ca chc sao m tng hp on
RNA mi to u 3'OH t do ca on mi. Enzyme DNA-polymerase III xc tc tng hp
mch b sung t u 3'OH t do ca mi, ko di mch. S tng hp cc mch n DNA
mi, v vy, ch i theo mt chiu xc nh t u 5' n u 3' (5'3').

S tng hp mch DNA mi xy ra mt cch lin tc trn mch khun c

chiu 3' 5' v gin on trn mch khun c chiu 5' 3'
Trn mch khun c chiu 3'5', mch mi c tng hp theo chiu 5'3' mt cch lin
tc, cng hng tho xon ca phn t DNA. on mi s c tng hp t im khi s
sao m. Khi xut hin u 3'OH t do ca on mi th enzyme DNA-polymerase III gn
vo v tng hp ngay mch b sung theo chiu 5'3' mt cch lin tc cho n im kt
thc sao m. Mch ny c tng hp nhanh hn nn ngi ta thng gi l mch nhanh
(leading strand). Nh vy, tng hp c si DNA mi b sung cho si lm khun ny,
enzyme DNA-polymerase ch cn c mt on mi

S
biu din cch sao chp gin on v lin tc trn 2 si DNA khun

Trn mch khun c chiu t u 5'3', vic tng hp mch mi phc tp hn, do qui lut
tng hp si DNA mi b sung lun thc hin theo chiu t u 5'3'. Mch mi b sung
cho mch khun ny c tng hp di dng tng on ngn gi l okazaki. Mi mt
okazaki c mt on mi. vi khun, di mt okazaki khong 1.000 n 2.000
nucleotide, cn t bo eucaryote, s lng nucleotide trong mt okazaki ngn hn so vi
vi khun. Hng di chuyn tng hp si mi ca DNA-polymerase III ngc vi
hng tho xon ca phn t DNA m.
Si con th hai ny c tng hp mt cch chm hn nn thng gi l mch chm hay
mch sau (lagging strand). Cc on RNA mi sau s b enzyme ribonuclease phn hu.
Cc l trng xut hin sau khi cc on mi mt i s c lp y nh hot ng ca

enzyme DNA-polymerase I. Cui cng, enzyme DNA-lygase ni cc lin kt phosphodiester

gia cc on, to nn mch DNA con hon chnh.
Ngoi chc nng tng hp si DNA mi theo chiu t u 5'3', enzyme DNA-polymerase
III cn c kh nng sa sai nh hot tnh exonuclease. Trn ng di chuyn tng hp,
nu n gp ch m nucleotide mi bt cp sai, n s li li ct b nucleotide sai v lp
nucleotide ng vo.
Kt qu, sau qu trnh sao m, hai phn t DNA con c hnh thnh c cu to ging y
nh phn t DNA m ban u, m bo thng tin di truyn c truyn t th h ny qua
th h khc mt cch chnh xc.

Sao m DNA si kp dng vng ca t bo procaryote

Sao m theo kiu hnh mt

S tng
hp DNA dng vng vi khun

DNA ca t bo procaryote thng c dng xon kp hnh trn. Qu trnh tng hp cng
c thc hin t mt im khi u (replication origine) gi l im xut pht v trin khai
ra c hai pha chc ba sao m v lan dn v hai pha, cui cng to ra hai phn t DNA lai.
C trng hp s tng hp ch xy ra v mt pha ca im khi u. Khi DNA dng vng
trn ang sao chp, quan st thy c dng hnh con mt (eye replication).
Mt n v sao m thng nht (t mt im xut pht) gi l replicon.
B gen ca cc sinh vt procaryote ch c mt replicon. Tc tng hp DNA ca E. Coli c
th t 50.000 nucleotide/pht, chu k sao m ko di khong 20 pht

Sao m kiu hnh trn xoay

Sa
o m theo kiu hnh trn xoay

Ngoi kiu sao m c dng hnh con mt nh trn, mt s vi khun v virus cn c kiu
tng hp khc gi l sao m hnh trn xoay. Qu trnh tng hp bt u bng s ct lin kt
phosphodiester ti mt im xc nh trn mt si DNA trn kp to ra hai u mt, mt u
kt thc bng nhm 3'OH v u kia l u 5'phosphat. S tng hp mt mch mi mt
cch lin tc bng cch ko di mch t u 3'OH ng thi dch chuyn theo dng xoay

trn, u 5'P c xoay ra ngoi v mt mch mi khc c tng hp gin on theo

okazaki, sau khi kt thc, hai phn t DNA con c hnh thnh

Sao m t bo eucaryote
t bo eucaryote c nhiu nhim sc th, a dng, mi nhim sc th l mt phn t DNA
nm lin kt vi protein, nn qu trnh sao m phc tp. Nhiu im sao m xy ra ng
thi, hay ni cch khc l trong cng mt thi im c nhiu n v sao m (replicon). V d
nh nm men bnh m S. Cerevisiae c 500 replicon.

Sao m t bo eucaryote

Qu trnh tng hp DNA t bo eucaryote phc tp hn v tc chm hn (3.000

nucleotide/pht).
c im quan trng trong qu trnh sao m t bo eucaryote l c nhiu n v sao m
xy ra, ng thi, trn mt phn t DNA v t bo c c ch kim sot nghim ngt qu
trnh ny. im no sao qua mt ln ri th khng lp li trc khi ton b phn t DNA
c ti to hon ton.
Sau khi sao m, cc DNA con c cu to ging y nh DNA m ban u c phn chia u
n v mi t bo con trong qu trnh phn bo.

Sa sai DNA trong t bo

Sa sai trong sao m
Sa cha nhng sai st trn DNA nhm khi phc li cu trc ban u l mt c tnh ca
mi t bo sng. Trong qu trnh sao m, cc baz nitt cng c th bt cp sai. S bt cp
sai thng xy ra l A bt cp sai vi C (A...C) v G bt cp sai vi T (G...T). S d c s
bt cp sai nh vy l do trng thi tn ti ca cc baz c nhng thay i, dn n s
nhn bit sai.
Nh c ch tng hp DNA lun lun khi ng t u 5 3, nn vic sao m c kim
sot v sa cha mt cch chnh xc. Cc enzyme DNA-polymerase I v III va lm chc
nng polymer ha va c hot tnh exonuclease theo hng 5 3 v 3 5. Nu trn
ng di chuyn bt gp mt nucleotide lp sai, chng s li li ct b v lp nucleotide
ng vo.
Khi h thng kim tra pht hin c sai st, cc enzyme endonuclease s ct b on sai,
sau , enzyme DNA-polymerase I s tng hp li cho ng v enzyme DNA-ligase s ni
li phc hi trng thi bnh thng. Khi tng hp nhn to phn t DNA (invitro) ngi ta

c tnh sai st l 105 , ngha l, trong 105 nucleotide c tng hp th c 1 nucleotide

sai. V d DNA trong t bo E. Coli c 3x106 nucleotide, nh vy c sau mi ln sao m th
c 30 nucleotide sai, hay cc nhim sc th ngi c 3x109 nucleotide, nh vy c sau
mi ln sao m th c 30.000 nucleotide sai. Nu vi tn xut sai ny th s lng t bin
s rt ln. Qua thc t nghin cu cho thy tn xut t bin trong cc qun th sinh vt l
nh hn rt nhiu.
Bng cch nh gi tn s t bin xut hin trong cc qun th, ngi ta c tnh s sai
st trong sao m nm trong khong 109. Tuy nhin, trong thc t, t l sai st cn thp hn
nhiu, vo khong 1010 n 1011, iu ny chng t s tn ti mt h thng kim tra v
sa sai rt hiu qu ca t bo.

Sa sai sau khi sao m

Mi trng sng v c th sng c mi quan h mt thit vi nhau. Cc bin ng ca mi
trng sng lun tc ng n cc sinh vt. Nhng thay i ca mi trng bn ngoi c
tc ng trc tip n b my di truyn, lm bin i n.

Cc tc nhn c th lm thng tn DNA trong qu trnh tn ti

- Cc tia v tr v cc tia phng x c nng lng cao c th lm bin i cc baz nit
nh gn thm cc nhm chc khc vo mch vng hay lm t vng, lm t cc lin kt
hydro gia hai mch hay lm ct mch ca DNA,...
- Cc tia cc tm trong nh sng mt tri thng gy nn s dimer ha cc baz thymine,
hnh thnh lin kt gia hai baz thymine nm k nhau, lm chng mt kh nng lin kt vi
baz adenine ca mch b sung.
- Tc ng ca cc thnh phn trong ni bo.
S tn thng phn t DNA c th gy ra do qu trnh trao i cht bt bnh thng gy ra
cc cht c hi v cc gc t do bt li, hay do hot ng ca cc enzyme khng ng b
lm tn ng nhng sn phm trung gian bt li.
Ngoi ra, cn c nhiu tc nhn ha hc khc ca mi trng bn ngoi cng gy nn s
bin i c th xy ra trn DNA.

Mt s kh nng gy bin i trn phn t DNA

- Gy mch hay t mch: Do si DNA rt mnh, bn thn n li xon cun nhiu ln nn
rt d b gy hay t mch do tc ng bn ngoi hay khi tho xon.
- Mt cc baz b sung, ngha l lm cho baz tng ng khng c cp (nh mt baz
purine).
- Bin baz nit ny thnh baz khc, gy nn s bt cp sai, nh: Khi mt nhm amin,
cytosine s bin n thnh uracil hoc 5-metyl-cytosine-desamin b nhn nhm l thymine.
- Cc baz nit c th tn ti di 2 dng ceton v enol nn dn dn bt cp sai.

- Gn thm nhm CH3 , C2H5: Khi gn thm nhm ankyl s lm thay i tnh cht ca
cc baz nit dn n bt cp sai.

C ch phng nga v sa sai

bo v v thch nghi vi nhng bin i, t bo c c ch phng nga v sa sai. Cc
c ch phng nga ca t bo nh h thng enzyme kh c, loi b cc c t, h thng
iu ho cn bng cn thit cho t bo, h thng enzyme tham gia sa sai. V d sau cho
thy h thng sa sai v phng nga ca t bo
- Enzyme Superoxide Dismutase (SOD) lm nhim v gim c: Enzyme SOD c pht
hin nm 1968, c m s EC.1.15.1.1 , c mt trong tt c cc t bo c chuyn ha oxy.
Chng xc tc phn ng phn hu gc superoxyt O
2

trong t bo theo s phn ng sau

2O
2

+ 2H+ H
2

O
2

+O
2

H
2

O
2

s c enzyme catalase hoc peroxydase chuyn ha. Do , SOD cng vi catalase c

xem l nhn t quan trng c th gii c, bo v v chng lo ha cho t bo. Gc O2 c
sinh ra lin tc v cng b phn hu khng ngng bi hot ng ca SOD, do , khi SOD c
hot cng cao th nng O
2

cng thp.

-H thng sa sai trong t bo rt a dng v c hiu qu vi s tham gia ca cc enzyme,

nh photolyase, DNA-metyl-transfersase, h thng enzyme m ha trong h thng gen
SOS.
vi khun E.Coli, do tc ng ca tia t ngoi lm xut hin cc thymine dimer (T=T) trn
DNA. Khi c nh sng, enzyme photolyase s c hot ha v ct b lin kt T=T, chuyn
v trng thi bnh thng v n nh.
H thng enzyme nuclease c th ct b ch sai hng bng nhiu cch v tng hp li cho
ng. Enzyme DNA-metyl-transferase c kh nng loi b gc metyl trong trng hp cc

baz b metyl ha. Tm li, t bo c c ch kim tra v sa nhng sai st nu c. Ngoi

ra, t bo cn c h thng SOS (cp cu): H thng ny hot ng khi t bo b tc ng
mnh bi cc tc nhn gy t bin, to nhiu sai hng trn DNA. Trong trng hp cp
bch c nhiu sai hng cn cp cu, cc gen SOS c m ra. Nu sa sai khng kp, t
bo phi chp nhn hoc b t bin hoc cht.
Trong t bo cn c h thng iu ho cn bng axit-baz, to iu kin thun li cho hot
ng trao i cht lm gim kh nng to cc cht c hi.
sinh vt a bo, vic kh c v loi b nhiu ha cht gy c c m bo bi gan
v thn.