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I.

CU TRC, VLDT CA VIRUS, VI KHUN: Vt liu di truyn vi khun l DNA mch kp vng trn Phn loi theo hnh dng: cu khun , trc khun, xon khun Vi khun thuc loi khng c nhn in hnh ( khng c mng nhn ). C quan cha thong tin di truyn l nhim sc th, n l phn t DNA di ,xon kp,khp kn, phn t DNA c trng lng 2 t Dalton, cha c 3000 gen,bao bc bi protein kim. DNA sao chp theo kiu bn bo tn. mt s vi khun c di truyn ngoi nhim sc th l cc loi plasmid transposon. Virus l nhng tc nhn gy nhim trng c kch thc nh nht (ng knh 20-300 nm)v trong b gen ca chng ch cha mt loi acid nucleic (RNA hoc DNA). Acid nucleic c bao bc trong lp v protein v bn ngoi cng c th c bao quanh mt mng lipid. Ton b phn t virus c gi l virion. Virus c b gen rt a dng: DNA : mch k p , mch n RNA: m ch kp, mach don B gen ca virus thng l mch thng hay dng vng trn Acid nucleic ca virus cha cc thng tin cn thit lp trnh cho t bo k ch b nhim bng tng hp cc i phn t c hiu cn cho s nhn ln ca virus. Trong chu k tng trng, mt s lng ln bn sao cc acid nucleic ca virus v protein v (coat protein) c to thnh. Cc protein v c tp hp li to v bao (capsid), v bao ny bao bc acid nucleic gip virus bn vng chng li mi trng ngoi bo v d dng xm nhp cc t bo cm th mi. C 3 dng tn ti ca virus bn trong t bo Viroid: Ch cha thnh phn RNA n tr n ,nh . c tnh truyn nhim n c. Virusoid: Ch cha thnh phn RNA khng c tnh truyn nhim n c. Virino:Ch cha cc thnh phn protein. vt liu di truyn ca virus: DNA: mt phn t DNA duy nht. nhiu hn 1 phn t DNA gi l h gen phn on. mt s loi khng c h gen hon chnh RNA Virus DNA: DNA c cu trc si n hoc kp, si thng hoc vng. Phn loi : DNA c kch thc ln (virus mn rp, zona ) DNA c kch thc nh ( virus bi lit, virus sinh khi u kh ) Virus RNA: RNA c cu trc mch n hoc kp. RNA dng mch n: mang ngha dng tnh hoc m tnh. RNA (+) c trnh t nucleotide trng vi trnh t nucleotid ca mRNA RNA (-) c trnh t b sung vi mRNA, do sn

phm c phin m t chnh h gen RNA ca virus m Retrovirus : vt liu di truyn RNA si n mang ngha dng tnh. h gen gm cc gen: gen gag, pol, env hai u gen c on trnh t lp tham gia vo qu trnh sao chp lng ghp v biu hin cc gen ca virus phage : virus ly nhim vi khun vt liu di truyn: DNA hay RNA si n hoc kp. Vd :phago M13 , T4 gy nhim Ecoli II. CU TRC NST CA TB EUKARYOTE 1. Cu trc NST: - cha mt phn t DNA duy nht c chiu di ln hn gp hng ngn ln so vi ng knh ca nhn (ng knh ca nhn khong 5m). V vy, ph hp th phi c s nn cht DNA trong nhim sc th. - Cc mc nn cht DNA: C bn mc nn cht phn t DNA. Xon kp DNA (c ng knh 2nm) khng b tc ng trong s nn cht ny. + Mc 1: DNA qun quanh li histon, to nn cc ht nucleosom c ng knh khong 10nm. Chromatin mang cc ht ny c thy ging nh mt chui ht, m mi ht l mt nucleosom v histon l phn li hay trc ca ht + Mc 2: Trong thc t, chromatin khng dng trng ht hay xu chui ko di (mc 1). Nhiu vng c ng knh khong 30nm trn cc si chromatin, do chui nucleosom t qun thnh si xon cht + Mc 3: si xon cht (mc 2) t xon thm mt ln na to thnh si siu xon c ng knh chng 200nm + Mc 4: s nn cht DNA ti a trong nhim sc th metaphase - Nhim sc th ca t bo nhn thc cn bao gm phc hp DNA-protein c t chc mt cch nh gn cho php mt s lng ln cc DNA c lu gi trong nhn ca t bo. S nn cht DNA c thc hin bng s qun cun DNA quanh cc li histon. Tiu n v ca nhim sc th l chromatin. n v c bn ca chromatin l nucleosome - Chromatin: l n v phn tch ca nhim sc th, chromatin phn nh cu trc ca nhim sc th nhng khng phi l duy nht cho bt k nhim sc th no. Chromatin chnh l phc hp ca 2 loi DNA-protein l: cc histon v cc protein khc histon. - Nucleosome: DNA qun quanh cc li histon, to nn cc nucleosom c ng knh khong 10nm

Ngay trc khi bt u phn chia (interphase), mi nhim sc th ca t bo nhn thc nhn i, to nn hai cu trc ging y nhau gi l chromatid; mi chromatid (=1/2 nhim sc th) cha mt bn thng tin di truyn y . Cc chromatid c ni lin nhau mt vng c bit gi l centromere, mi u ca mi chromatid c mt on nh gi l telomere. on gn vi centromere gi l heterochromatin v on gia heterochoromatin vi telomere gi l on euchromatin. 2. Vng telomere: Telomere: l mt khu vc ca cc chui DNA lp i lp li 2 u ca mt nhim sc th, bo v 2 u nhim sc th t s suy gim hay t cc phn ng tng hp vi cc nhim sc th ln cn. cc vng telomere ngn chn s lo ha ca gen khi gn kt thc ca nhim sc th bng cch rt ngn u kt thc ca nhim sc th, m nht thit phi xy trong qu trnh sao chp ca nhim sc th. Theo thi gian, do s phn chia t bo nn u kt thc telomere cng tr nn ngn hn. 3. Vng centromere: Centromere: l khu vc quan trng trong nhim sc th chu trch nhim l trung tm vn ng, l im trt ca nhim sc th trn dy t v sc i v cc cc trong phn bo. Centromere l ni phhaan ra bn tay: hai tay ngn (p) v hai tay di (q) ; cc tay xp thnh cp ging nhau. Centromere cha kinetochore, mt loi protein c bit lin quan trong s phn ly cc nhim sc th. Trnh t t centromere nm men plasmid: plasmid gim phn nh th n l NST eukaryote 2 loi chnh: Centromere im (~125 bp) & Centromere vng (4000-7000 bp) Ngn chn anaphase cho n khi NST gn vi t v sc c 2 cc III. MT S LOI TRNH T DNA EUKARYOTE: Ty theo mc hin din ca chng trong nhn, c cc loi trnh t DNA: Cc trnh t lp li nhiu ln. Vd: ng vt c v, cc trnh t chim 10 15% h gen. l nhng trnh t DNA ngn (10 200 kb), thng c sao chp mun, khng c phin m. khng m ha, thng tp trung nhng vng chuyn bit trn nhim sc th nh vng tm ng (trnh t CEN) hay u cc NST (trnh t TEL). chc nng ca cc trnh t ny cha r, c th chng tham gia vo qu trnh di chuyn DNA trn thoi v sc hay qu trnh sao chp ton vn ca phn DNA nm u mt NST. Cc trnh t c s ln lp li trung bnh. Vd: b gen ngi, cc trnh t ny chim 25 40%. Chng a dng hn v c kch thc ln hn ( 100 1000kb) cc trnh t lp li nhiu ln. cc trnh t ny phn b trn ton b b gen. chng l nhng trnh t khng m ha hay cng c th m ha cho rRNA, tRNA v RNA 5S. Cc trnh t duy nht: l cc gen m ha cho cc protein, c trnh t c trung cho protein.

IV.

MT S CU TRC CA DNR, RNA: 1. Cu trc kp tc (hairpin): Hairpin loop: (kp tc)

Cu trc hairpin l mt vng lp ca RNA thng tin (mRNA) c to ra khi mt si mRNA gp np v to thnh nhng cp base vi mt phn khc ca cng mt si. Cu trc trng ging nh mt vng lp hoc mt hnh dng U

Hairpin l mt loi ph bin ca cu trc th cp trong cc phn t RNA. RNA, cu trc th cp l hnh dng c bn ca trnh t cc nucleotide A, C, U, G to thnh sau khi c lin kt trong chui, chng hn gp hoc un li chui axit nucleic. dng hairpin mRNA c th c hnh thnh khi hai trnh t b sung trong mt phn t mRNA duy nht gp v kt hp vi nhau, sau s gp np ca phn t. Vng kp tc cng c th hnh thnh trong cc phn t DNA, nhng ph bin nht quan st trong mRNA.

C nhiu trng hp ca hin tng vng kp tc gia cc si nucleic acid. Mt v d v mt vng kp tc l trnh t ngng sao chp trong mt s prokaryote. Mt khi polymerase gp vng lp ny, n ri ra v kt thc phin m. Mt v d khc l RNA vn chuyn, trung tm trong tng hp protein, l mt phn c hnh thnh bi cc vng kp tc. Phn t RNA vn chuyn thc s c ba vng kp tc hnh thnh hnh dng c ba l. Mt trong nhng vng kp tc c cha mt chui gi l anticodon, nhn dng v gii m cc phn t mRNA ba nucleotide (mt codon) ti mt thi im trong qu trnh dch m. Cruciform (ch thp) Mt loi cu trc th cp ca DNA l Cruciform, c th c to thnh t s lp li mt cch o ngc khi cu trc hairpin hnh thnh trong mi chui si n.

1. cu trc hnh thnh ch thp l mt tnh nng DNA ph bin quan trng iu chnh cc qu trnh sinh hc. DNA c th tri qua qu trnh chuyn i to thnh mt hnh ch thp. cu trc ny quan trng i vi cc qu trnh sinh hc quan trng ca ti t hp DNA v sa cha xy ra trong t bo. DNA cruciforms ng mt vai tr quan trng trong quy nh cc qu trnh t nhin lin quan n DNA. Nhng cu trc ny c hnh thnh bng cch lp i lp li o ngc, v s n nh ca chng c tng cng bi supercoiling DNA. Cu trc hnh ch thp c v c bn quan trng i vi mt lot cc qu trnh sinh hc, bao gm c nhn rng, quy nh ca biu hin gen, cu trc nucleosome v ti t hp. n cng c lin quan n s tin ha v pht trin ca cc bnh nh ung th, hi chng Werner v nhng ngi khc. Cc cu trc DNA: DNA gm hai si polynucleotide i song song v b sung (trng hp ngoi l l cc phn t DNA si n trong mt vi virus). Hnh dng 3 chiu ca phn t c th khc nhau, tuy nhin n ty thuc vo cc iu kin trong DNA hin din v, trong mt s trng hp, ph thuc vo trnh t cc base. Cc cu trc ba chiu ca DNA m Watson v Crick m t c

gi l cu trc B-DNA. Cu trc ny tn ti khi nhiu nc xung quanh phn t v khng c trnh t base bt thng trong DNA iu kin hin din trong t bo. Cu trc B-DNA l cu hnh n nh nht cho mt trnh t ngu nhin ca cc nucleotide theo iu kin sinh l, v hu ht cc bng chng cho thy rng n l cu trc chim u th trong t bo. B-DNA l mt chui xon alpha, c ngha n l mt dng right-handed (hnh xon c, theo chiu kim ng h). N c khong 10 cp base (bp) cho mi 360- quay ca xon; cho nn mi cp base xon 36 so vi nhng base lin k. Cc cp base di 0,34nm , v vy mi vng quay hon chnh ca phn t l 3,4 nm. ng knh ca xon l 2 nm, v cc base vung gc vi trc di ca phn t DNA. Mt m hnh lm y khng gian cho thy B-DNA c mt c cu tng i mng v ko di. Xon c ca cc chui nucleotide to ra nhng rnh ln v nh trong xon, y l nhng c im quan trng cho s gn kt ca mt s protein kt hp vi DNA quy nh s biu hin ca thng tin di truyn. Mt s c im khc ca cu trc B-DNA cng vi nhng cu trc th cp khc tn ti trong nhng iu kin xc nh hoc trnh t base khc thng c trnh by trong bng sau:

Mt cu trc th cp c th gi nh l cu trc A-DNA - tn ti khi c t nc hin din. Cng ging nh B-DNA, A-DNA l mt xon alpha, nhng n ngn hn v rng hn so vi B-DNA v cc base ca n nghing so vi trc chnh ca phn t. C rt t bng chng cho thy DNA tn ti trong iu kin sinh l. Mt th cu trc hon ton khc c gi l Z-DNA, to thnh mt chui xon lefthanded. dng ny, khung ng - phosphate ngon ngoo qua li, cho nn c t tn l ZDNA (zigzag). Cu trc Z-DNA c th pht sinh trong iu kin sinh l khi nhng trnh t base c trng c mt. Cc b phn ca mt s gen hot ng to thnh Z-DNA, cho thy rng ZDNA c th ng mt vai tr quan trng trong iu phin m. V. DNA METHYL HA:
Metyl ha Metyl ha nhm mc ch bo v DNA ca cc on palindrome ngha l gn gc methyl (CH3) v tr cn thit nn khng b RE ct. v d e. coRI nhn bit chui

Khi c s metyl ha th enzyme khng nhn bit c v tr ct hn ch do DNA khng b ct nhng DNA ca phage khng c gn gc metyl s b ct bi RE. trong k thut gen enzyme metyl ha cn c gi l methylase . methylase c dng bo v on DNA cn gn vo Tt c cc chng e.coli u cha 2 enzyme metyl ha DNA l dam dcm dam (DNA adenine methylase): gn cc nhm methyl v tr N6 trong chui 5 GmeATC 3 Nhng DNA ca eukaryote khng b methyl ha v tr N6 ca adenine Dcm(DNA cytosine methylase): gn nhm methyl v tr C5 ca cytosine bn trong cc chui 5 CmeCAGG .. 3 hoc 5 CmeCTGG3 enzyme ch yu nh hng n s methyl ha ca dcm l EcoRII, enzyme BstNI c th nhn bit chnh xc cng mt trnh t nh EcoRII th DNA phi c chun b t cc chng E. coli dcm

1. Methyl ha: S bien oi histone: - Acetyl hoa histone (histone acetylation): nhom acetyl c gan vao nhom lysine tch ien dng trong uoi histone -> chromatin thao xoan -> phien ma - Methyl hoa( methylation) & Phosphoryl hoa: nhom methyl c gan vao chromatin. S gan them nhom photphate gan cac acid amin b ethyl hoa se lam chromatin thao xoan. S methyl hoa ADN: - mot so loai, s gan methyl vao cac nucleotide xac ng co the lam giam s phien ma. - S methyl hoa ADN co the lam gen bat hoat mot thi gian dai trong s biet hoa te bao - Trong s ghi dau bo gen (genomic impriting) s methyl hoa ieu hoa s bieu hien cua mot so gen xac nh nhan t cha hoac t me trong giai oan au cua s phat trien. 2. S in du S in du: cch duy tr hot ng ca gen T bo phn chia nhanh trong mt c th ang pht trin phi c cch duy tr kiu ni lng cc vng v hot ng ca gen to cho chng ha tnh thch hp vi loi m ca chng. C nhng bng chng chc chn rng kiu mu ca ng nhim sc cht v d nhim sc cht c th c truyn trc tip t mt t bo m n hai t bo con bng mt qu trnh gi l s in du (imprinting). Mt thnh phn ca h thng in du ny ng vt c xng sng l s methyl ha cytosin trong trnh t C-G nhng gen khng hot ng. S methyl ha c th chuyn cc kiu hot ng n cc t bo con : v ch c trnh t C-G c th b methyl ha bi enzim v v C-G lun lun bt cp vi G-C trn

methyl ha tn ti trn c hai si ca m : : :A:T:C:G:T:C:A: : : : : :T:A:G:C:A:G:T: : : m (Ch m ch cytosin b methyl ha bng cch thm vo mt nhm (CH3). S sao chp to ra mt cu trc lai nhng ni ADN b m b methyl ha m : : : A : T : C : G : T : C : A : : : si b m : : : T : A : G : C : A : G : T : : : si mi m Mt loi enzim c bit l methylaz tc dng vo ADN c cytosin b methyl ha trong trnh t C-G v methyl ha cytosin tng ng trn si mi. Nh vy, cch thc ca s methyl ha (s bt hot ca gen) c truyn li nguyn vn. Tuy nhin, nu c hai nhm methyl ti mt v tr tnh c b mt, hot ng ca gen b bin i t bo con. C bng chng chc chn rng mt s vn c lin quan n s lo ha l do s methyl ha mt dn theo thi gian v t dn ti s hot ha ca nhng gen khng thch hp trong t bo. S kh methyl ha trong cc giao t c th truyn kiu hot ha gen sai n th h con v ngy nay nhiu bnh c xem l do cc sai st trong s in du ny. DNA METHYLATION (http://www.premierbiosoft.com/molecular_beacons/dnamethylation/index.html) Gii thiu v methyl ha DNA Sau mi chu k sao chp DNA, sa i mt s xy ra trong DNA. Methyl ha DNA l mt sa i bi tng hp nh vy.Methyl ha DNA c chng minh bi nghin cu c th hin trong mt s qu trnh sinh hc nh quy nh ca cc gen in du, bt hot nhim sc th X, v gen c ch khi u im lng trong cc t bo ung th. N cng hot ng nh mt c ch bo v c thng qua bi DNA mm bnh (ch yu l vi khun chng li hot tnh endonuclease ph hy bt c DNA nc ngoi. C ch DNA methyl ha Trong DNA, methyl ha thng xy ra cc o CpG, mt khu vc giu CG, thng ngun ca vng promoter. Ch "p" y c ngha rng C v G c ni vi nhau bng mt lin kt phosphodiester. ngi, methyl ha DNA c thc hin bi mt nhm cc enzyme gi l methyltransferases DNA. Nhng enzyme ny khng ch xc nh cc m hnh methyl ha DNA trong qu trnh pht trin ban u, nhng cng chu trch nhim cho vic sao chp nhng m hnh ny cc si c to ra t sao chp DNA.

mt si ADN:

khc

nn

kiu

MethyLight kho nghim thit k h s methyl ha DNA MethyLight kho nghim, mt thit b thm d da trn TaqMan methyl ha h s k thut, c th c s dng c hiu qu h s methyl ha DNA, methyl ha DNA o hoc pht hin cc alen methyl ha trong s hin din ca nhng ngi unmethylated. Do cc c trng ca TaqMan xt nghim da trn, n l nhiu hn chnh xc hn xt nghim pht hin im cui nh methyl c th PCR (MSP). Cc mu DNA l ln u tin c iu tr vi sodium bisulphite chuyn i cc c s cytosine unmethylated uracil. Cc cytosines methyl ha, mt khc, vn khng b nh hng. Mi Methyl c th c thit k vi DNA chuyn i bisulphite. Mu ny c khuch i bi PCR thay th uracil vi thymine. Mt TaqMan thm d sau c s dng pht hin cc si methyl ha. Tnh nng ni bt h tr cho cc xt nghim MethyLight trong thit k Beacon a) D on ca cc o CpG trong chui DNA. b) Thit k methyl mi c th v thit b thm d TaqMan . c) Thit k mi kim sot ph hp v TaqMan thm d cc chui tng ng unmethylated v khng c iu tr. d) Phn tch mi pre-designed/published v / hoc TaqMan u d v thit k ca oligos tng thch. e) mi v TaqMan u d c thit k bng cch trnh cc khu vc tng ng c xc nh bng cch t ng gii thch cc kt qu tm kim BLAST. VI. YU T CHUYN V (TRANSPOSABLE ELEMENTS) Mt s khi nim chung: -Tranposon : mt on AND c kh nng c chuyn v. -LINEs ( Long interspersed elements) : on DNA c chiu di t vi trm cho n 9000 cp base. -SINEs ( Short interspersed elements) : on DNA ngn khong 100-400 cp base i din cho phn t RNA phin m ngc. c im chung: - Thng i km xen on, mt on v cu trc li b gene. - H thng ti t hp gene c th tc ng. - C kh nng gy ra t bin xu dn n ung th v ph hy gene. Chuyn v: C 3 cch chuyn v: - Chuyn v khng sao chp: phn t chuyn v c ct ra DNA cho v gn vo DNA mc tiu . V d nh: Tn10, Tn5, Tn7. - Chuyn v sao chp: DNA nhn i v bn sao gn vo v tr mi to cointergrade(cng gn). V d: Tn3, Mu. - Retrotransposition: qua trung gian RNA, to ra cDNA v xen cDNA vo v tr mi. ngha :

- Thng gy ra t bin nn c th l ngun nguyn liu cho tin ha. - Mt s trng hp c li cho t bo ch : + Telemorase: lin quan n s phin m ngc ca LINEs. + RAG-1 v RAG-2 cn cho vic kt hp khng th v th th t bo T. C ch ny c s gp phn ca chuyn v khng sao chp. + To nn kh nng khang DDT cn trng. Ngun tham kho: DI TRUYN HC . Phm Thnh H. [501-505] 8/2010 http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/T/Transposons.html , 6/12/2011 accessed time.
VII. YU T CHUYN V (TRANSPOSABLE ELEMENTS)

Mt s khi nim chung:


Cc phn t chuyn v (transposable elements) hay phn t di ng (mobile elements) l nhng trnh t ADN c kh nng di chuyn t v tr ny n v tr khc (mc tiu) ca b gen (cng b gen hay khc). S di chuyn ADN ny c gi l transposition (s chuyn v).

-Tranposon : mt on AND c kh nng c chuyn v. -LINEs ( Long interspersed elements) : on DNA c chiu di t vi trm cho n 9000 cp base. -SINEs ( Short interspersed elements) : on DNA ngn khong 100-400 cp base i din cho phn t RNA phin m ngc. c im chung: - Thng i km xen on, mt on v cu trc li b gene. - H thng ti t hp gene c th tc ng. - C kh nng gy ra t bin xu dn n ung th v ph hy gene. Chuyn v:
Tranposition gm cc kiu chnh: IS:trnh t xen on (insertion sequence) l cc transposon ngn. Transposon (k hiu Tn): Cc phn t di ngdi(khong 5000 bp) c cha mt hoc vi gen. N thc hin: + Transposition khng sao chp (nonreplicative): Phn t chuyn v c ct khi ADN cho (donor) v c gn vo phn t ADN mc tiu. Thuc loi ny c Tn10, Tn5, Tn7. + Transposition sao chp (replicative): ADN c nhn i v bn sao c xen vo v tr mi to cointergrat (cng gn) v c Tn3, Mu. Retrotransposition: S di chuyn qua trung gian ARN, nh reverse transcriptaz to thnh cADN, v s xen on cADN vo v tr mi.

ngha : - Thng gy ra t bin nn c th l ngun nguyn liu cho tin ha. - Mt s trng hp c li cho t bo ch : + Telemorase: lin quan n s phin m ngc ca LINEs. + RAG-1 v RAG-2 cn cho vic kt hp khng th v th th t bo T. C ch ny c s gp phn ca chuyn v khng sao chp. + To nn kh nng khang DDT cn trng.

Vai tr trong vic kim sot cht d nhim sc v biu sinh

Ngun tham kho: DI TRUYN HC . Phm Thnh H. [501-505] 8/2010 http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/T/Transposons.html , 6/12/2011 accessed time.

Cc trnh t xen on IS
Cc transposon n gin nht l cc trnh t xen on (insertion sequence) v c k hiu bi tip u ng IS km s th t nh IS4. Cc phn t IS l cc cu phn bnh thng ca ADN vi khun v plasmid. Dng E.coli chun thng cha vi bn sao (<10) ca bt k mt trong cc IS chung thng gp. m t s xen on vo im c bit, k hiu 2 chm kp c s dng nh :: IS1 ch IS1 xen vo phage . Cc IS l nhng n v t tr, mi mt trong chng m ha cho ch mt protein cn thit cho s chuyn v bn thn chng. Trnh t ca mi loi IS c khc nhau, nhng trong t chc cu to c nhiu tnh cht chung. gia c gen transposaz tnp, hai u c lp on o ngc (IR Inverted repeats). S xen on ca IS tiu

im (target) c minh ha trn hnh 27. Cu trc transposon v s chn vo ADN mc tiu nu trn hnh
28. Cc u mt ca transposon c trnh t lp li o ngc (inverted repeat - IR). Trong v d ny, tiu im c 5 bp. Cc u mt ca transposon gm cc lp on o ngc 9 bp c nh s t 1 n 9. IRIRDNA mc tiu

Hnh 1: Cu trc IS v s chn vo ADN mc tiu

hai u ca IS lun c hai trnh t lp on trc tip (direct repeat) ngn. Cc trnh t ny dao ng ty transposon, nhng c nh i vi mi loi IS. Chiu di ca phn ln lp on trc tip l 9, chng xc nh cc u mt ca transposon.

Mc tiu

Hnh 2: S cu trc ca transposon c cc IS, cc lp on o ngc IR (inverted repeat) v to lp on trc tip hai u ADN im mc tiu

Transposon khng sao chp v sao chp

S chuyn v ADN theo 2 kiu khc nhau (sao chp v khng sao chp hay bo tn (conservative)) v c nhng tnh cht chung. S din bin transposition sao chp (transposon Tn3) c nu trn cc hnh sau: ct v ni, s cng gn v phn tch.
Hnh 3: S v transposition sao chp v khng sao chp

Hnh 4: S transposition sao chp (Tn3)

Tip theo l s cng gn (Cointegrat Formation) 2 loi ADN

Hnh 5: Cointegrat) ca 2 loi ADN


Cui cng l s phn tch (Resolution) ra 2 loi ADN

Hnh 6: S phn tch ra 2 loi ADN


Qu trnh transposition c nhng c im sau y: C hai u ca phn t mang gn nh mt trnh t c nh hng o ngc nhau. Cc transposon m ha t nht mt protein l transposaz. Transposaz gn c hiu v ct cc trnh t cui thc hin chuyn v. Cc transposon to bn sao ngn ( 12bp) ca ADN im mc tiu trong transposition. Chiu di ca trnh t ny c trng v khng i i vi mt phn t nht nh v c to ra nh s ct so le (staggered cleavage) ca ADN mc tiu nh transposaz. C ch transposition c nghin cu chi tit phage Mu v c th tm tt nh sau: S nhn bit v bt cp 2 u mt ca transposon nh transposaz hnh thnh cu trc chuyn bit protein - ADN. S hnh thnh phc hp c kch thch bi cc protein HU vIHF l cc nhn t tham gia ti t hp im chuyn bit Mu transposaz, ging nh Int protein, c 2 vng gn c lp. S ct khc (nicking) do transposaz to ra 3-OH mi u ca Mu. Ct tiu im nh transposaz to u mt so le 5 - P di 5 bp.

S ni u 5 - P ca tiu im vi 3-OH ca transposon to cu trc trung gian chuyn mch (strand transfer intermediate).

Tn10 c l s dng c ch tng t thc hin transposition khng sao chp hay bo tn (conservative
transposition).

S chuyn v ngc (Retrotransposition)


Nhm cc transposon, c pht hin nm men (Ty) v Drosophila (copia) v c lin quan n retrovirut, di chuyn n v tr mi qua trung gian ARN. Transposon ca nm men Ty, ging vi retrovirut, c lp on cui

di (long terminal repeat) c gi l trnh t nm 2 pha on m ha. Cc protein c m ha, c trnh t tng ng vi integraz ca retrovirut v reverse transcriptaz, rt cn thit cho transposition.
Phng thc phin m ngc ca cc phn t di ng tng t nh ca retrovirut. C s tng t v cu trc gia retrovirut v cc retrotransposon nm men (Ty) v Drosophila(copia). S chuyn v ngc to bn sao ca phn t v tr mi, trong khi phn t cho ban u vn gi nguyn cu trc khng bin i. Do vy, retrotransposition to nn mt t t on v cc ti cu trc (rearrangement) ca b gen t bo ch. Nhng bin i ca b gen lin quan vi retrotransposition s dn n vic lm ngng hay hot ha cc gen, m mt s c th gy ung th.

Cc transposon gp phn vo s tin ha ca plasmid, v vi chng c cho thy cc plasmid R c c tnh khng khng sinh thng qua cc transposon.
Hnh 7: Transposition khng sao chp hay bo tn conservative transposition)

Mi tn ch cc im ct v sau ni li theo trt t mi.

Gy t bin bng transposon


Cc ng dng ca transposon bao gm: Gy t bin (mutagensis) bng transposition.
Hnh 8: Transposon Tn10 chn vo A lm mt chc nng,

nhng khng tetra.

Cng c cho k thut di truyn in vivo (in vivo Gentic Engineering). Nhn t trung gian khng khng sinh (mediator of antibiotic resistance) y cp ch yu n gy t bin nh transposon (Transposon Mutagensis). S chn transposon vo gia gen s dn n t bin lm mt chc nng bnh thng ca gen (hnh trn). Transposon cung cp mt phng tin d dng to cc t bin trn Nhim sc th. Mt thun li cho gy t bin bng transposon l n c cha 1 gen khng khng sinh, m gen ny dng lm marker (du chun) nhn bit transposon c chn vao. Trc tin cc dng mang gen khng khng sinh c phn lp t mi trng giu dinh dng, ni cc t bin khuyt dng tng trng to khun lc. Sau chng c th c sng lc trn mi trng gii hn c cung cp cc cht tng trng no xc nh cht no cn thit cho t bin. Cc transposon cng hu ch cho s chn 1 maker gen khuyt dng (auxotrophic) vo trong dng sinh vt hoang di. Thng th cc t bin khuyt dng kh phn lp trc tip. Nhng nu marker khuyt

dng c a vo cng transposon vi marker khng khng sinh, th c th qua chn dng khng khng sinh m xc nh marker khuyt dng. Hai transposon thng c s dng trong gy t bin l Tn5 (khng neomycin v kanamycin) v Tn10 (cha 1 marker khng tetracycline). Integron l cc transposon c th thu nhn v biu hin cc gen t nhiu ngun khc nhau. Tuy nhin, khc
vi cc transposon c chn vo mt cch ngu nhin, cc integron chn vo c tnh chn lc cao (high selective) trong cc im xen on (insertion site) ca chng, thng th c chn vo trong plasmid.

Integron cha gen m ha cho protein c gi l integraz, cn thit cho s ti t hp im chuyn bit (site-specific recombination). B gen ca phage (lambda) chn vo Nhim sc th ca E. coli mt v tr c bit nh hat tnh ca enzym integraz . Integron cng cha trnh t ADN c bit cho php integraz chn cc nhm gen gi l cassette vo Nhim sc th, m trn c promoter cho php cc gen cassette mi c
biu hin.