Các Vector Sử Dụng Để Chuyển Gen ở Thực Vật

Cc vector s dng chuyn gen thc vt

Bi:

Trn Quc Dung

Cc vector bin np vo t bo thc vt s dng Agrobacterium

S gy ra cc khi u do Agrobacterium

Agrobacterium tumefaciens v A. rhizogenes l hai loi vi khun sng trong t gy rabnh khi u hnh chp (crown gall) (Hnh 1v hnh 2 ) v bnh lng r (hairy root) ccv tr tn thng ca thc vt hai l mm.

Ch rt t thc vt mt l mm thuc h Liliaceae v Amaryllidaceae l d b bnh khiu hnh chp. L do gii hn vt ch ny hin nay cn cha c bit. Mt ln khi u,s sinh trng khi u c th tip din khi vng mt vi khun v m khi u c th csinh trng v trng bng nui cy m trong cc mi trng thiu s b sung auxinv cytokinin ngoi sinh m bnh thng l cn thit xc tin s sinh trng ca mthc vt in vitro. M khi u tng hp amino acid mi v cc dn xut ca ng cbit chung l opine. Loi opine tng hp trong khi u (v d nh nopaline, octopine,agrocinopine, mannopine v agropine) ph thuc vo dng Agrobacterium khi u shnh thnh khi u. Octopine v nopaline l hai loi opine c ngun gc t arginine vd dng pht hin nht trong m khi u hnh chp. Do nhiu dng Agrobacteriumtumefacien ph bin c thit k theo kiu octopine hoc nopaline. Agropine, mt dnxut ca ng c tm thy ph bin trong cc khi u lng r gy ra bi A. rhizogenes.Agrobacterium chu trch nhim i vi s hnh thnh khi u d ha mt cch c chnlc opine m n tng hp c v s dng opine nh l mt ngun cacbon v nit.

C vic gy ra khi u v tng hp opine lin quan vi s c mt ca mt loi plasmidc kch thc ln l Ti-plasmid (Tumour inducing =Ti= gy ra khi u) trong t bovi khun A. tumefaciens v Ri-plasmid (Root inducing = Ri = gy ra lng t) A.rhizogenes.


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Bnh khi u hnh chp cy mm xi xi (Rubus) do A.tumefaciens gy ra

Cc khi u hnh chp cnh to

S cu trc t bo vi khun Agrobacterium tumefaciens

Ti-plasmid ca Agrobacterium tumefaciens

Ti-plasmid c tm thy trong tt c cc dng A.tumefaciens gy c, c kch thckhong 200-250 kb. Chng c duy tr n nh trong Agrobacterium nhit di30oC. Bng phng php lai DNA-DNA v lp bn chui kp d hp (heteroduplexmapping), ngi ta xc nh c Ti-plasmid c 4 vng tng ng. Kt qu phntch di truyn cho thy vng T-DNA (transferred DNA) v vng gy c (virulence) linquan n s hnh thnh khi u trong khi hai vng khc lin quan n s tip hp v sti bn ca plasmid trong Agrobacterium.

Trong khi hnh thnh khi u, T-DNA c chuyn vo t bo thc vt v hp nht vigenome nhn. T-DNA n nh trong genome nhn. Lai Ti-plasmid vi DNA ca khiu cho thy T-DNA trong t bo thc vt l tng ng song song vi T-DNA trongTi-plasmid ca Agrobacterium. Kt qu ny chng t khng c s sp xp li v tr ca


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T-DNA trong lc khi u c to thnh. Mt hoc nhiu bn sao ca T-DNA c th cmt cc on lp ni tip. Chng cng c th tch ra v lin kt vi cc vng khcnhau ca DNA thc vt. V tr hp nht ca T-DNA vo DNA thc vt l hon ton ngunhin. Cc vng tng ng vi T-DNA c tm thy cc Ti-plasmid khc nhau(Hnh 1.21B ). Trong cc dng A.tumefaciens kiu nopaline c s dng ph bin.Vng T-DNA c kch thc khong 24 kb. mt s khi u hnh chp kiu octopine,hai on khng k nhau c tm thy l TL v TR. TL c kch thc 14 kb, c mttrong tt c cc dng t bo bin np v c chc nng tng ng vi T-DNA cc dng t bo nopaline. TR c kch thc 7 kb, c hnh thnh t pha bn phi caTL-DNA trong Ti- plasmid, khng pht hin thy trong tt c cc dng khi u nhngkhi s bn sao ca n khc vi TL ngi ta gi thit l xy ra mt qu trnh chuync lp.

T-DNA c phin m trong cc t bo khi u (Hnh 4B), to ra nhiu mRNApolyadenyl. Cc loi sn phm phin m ca T-DNA tch ly li l tng i thp sovi cc mRNA ca thc vt khc. Gii trnh t T-DNA kiu nopaline cho thy c 13khung c m ln (open-reading frame) trong khi TL-DNA v TR-DNA kiu octopinetng ng l 8 v 6. Cc sn phm phin m pha cnh phi ca T-DNA kiu nopalinec chc nng tng ng vi cc sn phm phin m pha cnh phi ca TL-DNA(Hnh4B). Ton b s t chc ca cc gen trn T-DNA v cc vng ln cn l tng tvi cc gen ca genome eukaryote mc d chng khng cha intron. So snh cc trnht, s pht sinh t bin mt on v gen nhy cng nh s tng sinh ca cc sn phmgen ring l E.coli c s dng pht hin ra chc nng mt s sn phm genc m ha bi T-DNA. Mt gen trong vng TL octopine m ha enzyme octopinesynthase. Ti-plasmid nopaline, cc gen opine synthase bao gm nopaline synthase(nos) v agrocinopine synthase (acs). Trong Ti-plasmid octopine, vng TR m ha haiprotein chu trch nhim i vi s tng hp manopine v mt sn phm ca gen xctc cho s bin i ngc manopine thnh agropine. Locus tmr m ha mt enzymelin quan n h thng cytokinin v s t bin y dn n s tng sinh r t cckhi u hnh chp gy ra mt s loi (cc th t bin r). Cc locus tms 1 v tms 2lin quan n s tng hp khng iu ha ca auxin v s t bin mt trong hai locusgy nn s tng sinh chi t cc khi u hnh chp nhiu loi thc vt (cc th tbin chi). V vy, T-DNA mang cc gen (tms 1, tms 2 v tmr) m sn phm ca chngkhng chu s iu ha bnh thng ca cc con ng chuyn ha thc vt lin quann s tng hp phytohormone v iu ny gy ra kiu hnh ung th. Do cc gen tms1, tms 2 v tmr c xem l cc gen ung th. Tuy nhin cn lu rng cc gen c mha bi T-DNA khng i hi phi chuyn T-DNA vo t bo thc vt v cng khngduy tr s n nh ca n trong genome thc vt.

Ri-plasmid ca Agrobacterium rhizogenes

S khi u ca bnh lng t do A.rhizogenes tng t nh s bin np nhA.tumefaciens. Di s kim sot ca vng gy c, hai vng phn tn ca Ri-plasmid


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c chuyn vo genome thc vt (Huffman, 1984; De Paolis, 1985). Cc vng nyc tn l TL (T-left T-DNA) v TR (T-right T-DNA). TR T-DNA cha cc gen snxut opine (mannopine hoc agropine) v cc dng c nh r c im nh ccgen opine c trng ca chng (De Paolis, 1985). Mt khc TR T-DNA cn cha haigen m ha cho s tng hp auxin. Cc gen ny tng ng rt cao vi cc gen auxinca A.tumefaciens v cho thy c th b sung thm cc dng mang cc gen auxint bin ca chng (Offringa, 1986). TL T-DNA khng tng ng vi T-DNA caA.tumefaciens (Huffman, 1984). Ton b TL T-DNA ca c gii trnh t v ncha ti thiu 11 khung c m, tng t vi cc khung c m c tm thy genome ca eukaryote. Chng c promoter v cc yu t polyadenine ha cn thit thc hin chc nng khi chuyn vo trong genome thc vt (Simpson, 1986). Cc genny khng tng ng vi bt k gen no.

Cu trc v s biu hin ca Ti-plasmid kiu octopine v nopaline

A. Bn ca Ti-plasmid kiu octopine (pTiAch5) v kiu nopaline (pTiC58) cho thyv tr tng i v kch thc ca cc vng chc nng chnh.

Cc v tr t m ch cc vng tng ng ln ca 2 plasmid.

//: Cc on lp trc tip 25 bp; CON: vng m ha chc nng tip hp


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ORI: vng m ha chc nng ti bn v khi im ti bn

B. Bn ca vng T kiu nopaline v octopine cho thy cc vng chc nng chnh,cc vng tng ng v cc sn phm phin m.

Cc sn phm phin m c bit l: 3(nos): nopaline synthase; 3 (ocs): octonopalinesynthase; acs: agrocinopine synthase; 1(tms 1): tryptophan mono-oxygenase; 2(tms 2):indole-3-acetamide hydrolase; 4(tmr): DMA transferase; agr: 3 sn phm phin m cncho s tng hp agropine.

bit v cng khng c s tng ng c ngha no i vi T-DNA (kiu octopine)ca A.tumefacien. TR T-DNA l khng cn thit i vi s duy tr kiu hnh lng t.Tuy nhin nhiu loi, cc dng Agrobacterium c c TL v TR T-DNA l c nhiuhn so vi cc dng ch mang mt T-DNA (Vilaine v Case-Delbart, 1987).

C ch chuyn T-DNA

Cc vng T-DNA ca c A.tumefaciens v A.rhizogenes u nm bn cnh ccon lp trc tip 25 bp (Hnh 1.11) v cc im kt thc ca T-DNA hp nhttrong genome thc vt nm st vi cc trnh t ny. Trnh t lin ng ca bin T-DNA l GGCAGGATATTC/GA/G GT/GTCTAAA/TT/C. Hu ht cc nghin cuc ch chuyn T-DNA t Agrobacterium vo t bo thc vt c tin hnh A.tumefaciens. Vic loi b b phi ca Ti-plasmid kiu nopaline lm mt s hnhthnh khi u, nhng khi on oligonucleotid tng ng vi b phi c to dng vis nh hng chnh xc vi Ti-plasmid thiu b phi th s hnh thnh khi u li cphc hi (Wang, 1984), cho thy rng cc trnh t lp 25 bp phn cc v tc ng u(cis-acting).

S tip xc ca Agrobacterium vi cc hp cht gii phng t m thc vt b tn thng lm cho vng vir ca Ti-plasmid phin m (Hnh 1.22). Trong qu trnh ny mt chtc hot tnh ha hc cao v c trng c nhn bit l acetosyringone. Gen vir Av gen vir C c biu hin vi khun sinh trng sinh dng mc d gen vir C chc phin m mc thp. Khi Agrobacterium gp dch r ca cc t bo thc vt btn thng, hoc acetosyringone tinh khit, sn phm ca gen vir A (c th lin kt vimng) s nhn din v tng tc vi acetosyringone v truyn tn hiu ngoi bo votrong t bo ny lm hot ha sn phm ca gen vir C. Sau protein vir C bin ihot ha lm cho cc gen vir B, C, D v E khng hot ng v lm tng cng s phinm ca gen vir C.

S cm ng gen vir xy ra nh s xut hin cc im t si n trong cc trnh t bin25 bp nm mp ca T-DNA (Stachel v Zambryski, 1986; Albright, 1987) v s xuthin phn t si n mch thng tng ng vi T-DNA . Cc sn phm ca operon virD c hot tnh endonuclease (Yanofsky, 1986). Bng mt c ch tng t s tip hp


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ca vi khun, T-DNA c chuyn sang t bo thc vt v xen mt cch n nh voDNA nhn (Stachel v Zambryski, 1986). S kin ny c th lin quan n protein cm ha bi operon vir E (Winans, 1987). M hnh m t cc s kin xy ra mc phnt trong s tng tc gia t bo thc vt v Agrobacterium hnh thnh khi u hnhchp c trnh by hnh 5.

Plasmid Agrobacterium l vector bin np

Kh nng chuyn mt cch t nhin cc trnh t DNA xc nh vo genome thc vtca Agrobacterium c s dng vo vic pht trin cc vector bin np thc vt.Cc vector ny li dng mt s c tnh bm sinh ca Agrobacterium l qu trnh binnp trung gian (mediated transformation process). Vo u thp nin 1980, mt s nhmnghin cu Ti-plasmid loi b tt c cc gen onc ca T-DNA. Khm ph quan trngth nht l cc gen onc ny khng cn thit i vi vic chuyn T-DNA vo t bo thcvt v khng hp nht vo DNA nhn. V vy cc gen ny c th c thay th. Nkhng ch cho php xen DNA ngoi lai vo m cn cho php loi b cc chc nng cagen onc. Tuy nhin cn lu rng, nos hoc ocs l cc gen marker hu ch i vi sbin np bi v hot tnh enzyme ca cc sn phm gen ca chng c th c pht hinbng mt xt nghim n gin. Cho n nay, gii hn kch thc ca on DNA xenvo cha c cng b. S kin quan trng th ba l s khm ph cc sn phm cagen vir cn hot ng chc nng trong trans. Cui cng, T-DNA khng gy ung th cmt trong ton b thc vt ti sinh c truyn li cho th h sau theo kiu di truynMendel.

Cc thnh phn c bn ca vector Ti-plasmid khng gy ung th

c im ca chuyn gen qua trung gian Agrobacterium l bt k DNA no todng u c th c chuyn vo genome ca t bo thc vt hai l mm. DNA ngoilai phi c nm mp cc trnh t bin ca T-DNA v duy tr n nh trong dngAgrobacterium mang nhm b sung y ca cc gen vir dng cis hoc trans (nh vtrn plasmid h tr gy c phn tn). Cc gen trn nhim sc th ca Agrobacteriumkt hp vi vng gy c c m t v c lin quan vi vic vi khun nhn ra mtthnh phn ca b mt t bo thc vt, ri gn vo sau .


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S tng tc gia Agrobacterium vi t bo thc vt v c ch chuyn T-DNA

nhn ra cc t bo c bin np, cc gen tri khng thuc khng sinh ca vikhun c xen vo trong cc trnh t lp 25 bp di s kim sot ca T-DNApromoter v cc tn hiu polyadenyl ha. Cc gen khng thuc khng sinh dng khmnh th biu hin mt cch c hiu qu trong t bo thc vt bt k mi trng ditruyn no. Vic lin kt mt cch cht ch cc gen marker vi DNA ngoi lai c 2 lich: th nht l chn lc trc tip cc m thc vt c bin np DNA ngoi laivo mt cch chc chn; th hai l m bo DNA ngoi lai trong cc dng bin np ctrng chn lc khng xen vo vng genome khng c phin m.


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Cc vector bin np thc vt khng gy ung th da trn Ti-plasmid

C A.tumefaciens v A.rhizogenes u c s dng chuyn DNA ngoi lai vo tbo thc vt. Nhiu vector bin np da trn Ti-plasmid c pht trin (Bng 1.1 vbng 1.2). Cc vector ny khng cha bt k trnh t ung th no v v vy t bo thcvt c th sinh trng bnh thng sau khi chuyn DNA vo nhn ca n. Cc vectorkhng gy ung th hin ang s dng c th c chia lm hai loi l cis v trans, davo vic c hay khng cc vng T-DNA nm mp cc trnh t lp trc tip 25 bp trncng n v ti bn (replicon) nh cc gen vir hoc trn mt plasmid phn tn (Hnh1.13). Trc y, loi vector thng c cp n l vector lin hp (co-intergrativevector), tuy nhin gn y vector nh th (binary vector) l ph bin hn.

Cis vector

y l cc vector c ngun gc t Ti-plasmid kiu di m gen onc trn T-DNA cloi b v trong mt s trng hp c thay th bng mt on DNA c hiu cvng tng ng vi mt vector to dng nh m ch c th ti bn E.coli. Chin lcvector ny ph thuc vo s lin hp trong A.tumefaciens gia cc vng tng ngtrn Ti-plasmid sa i (h tr mang gen vir) v mt vector to dng nh E.coli(vector trung gian) mang gen marker chn lc s hot ng chc nng trong cc t bothc vt v cc trnh t duy nht cho vic xen DNA ngoi lai vo.

Vector trung gian mang trnh t DNA ngoi lai thng c a vo A.tumefaciensbng s tip hp v s dng s chn lc thch hp s thu c th nhn tip hp viDNA ngoi lai n nh trong T-DNA l kt qu ca ti t hp tng ng (Hnh 6A).


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S h thng vector lin hp (A) v vector nh th (B)

VIR: vng gy c; HOM: cc vng tng ng trong s ti bn c th xy ra ivi s lin hp; LB: bin tri; RB: bin phi; MCS (multicloning site): cc trnh t todng; PTM: marker bin np thc vt; RES: marker khng thuc khng sinh chn lci vi s c mt ca cc trnh t vector trong vi khun ch; oriT: khi im chuyn;ColE1: khi im ti bn t plasmid ColE1; RK2: vng c ph khi im ti bn rng,c ngun gc t pKR2

mt s vector nh pGV3850 (Hnh di), c hai bin ca T-DNA l trn Ti-plasmid sa i. Trong cc vector nh pGV2260 (Debleare, 1982), cc trnh t lp 25 bp l trn vector trung gian, trong khi vector pTiBS3-SE (Fraley, 1985), bin phi l trnvector trung gian v bin tri trn gen vir ca plasmid. Bt k u trong cc v trkhi u ca cc trnh t lp 25 bp, kt qu thc sau khi lin hp l s xen cc trnht DNA ngoi lai bin T-DNA lp li trn Ti-plasmid sa i. V vy cc dngA.tumefaciens mang cu trc ny, T-DNA c chuyn vo genome thc vt trong suts bin np l kt qu hot ng ca vng vir dng cis.


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Cu trc v s s dng vector lin hp pGV3850

Bn cho thy cu trc ca vector bin np thc vt pGV3850 v vector trunggian pGV1103 v kt qu ca vic hp nht pGV1103 vo pGV3850. LB: bin tri;RB: bin phi; Pnos: promoter nopaline synthetase; npt-II: trnh t m ha neomycinphosphotransferase-II t Tn5; ocA: trnh t polyadenyl ha octopine synthase; E:EcoRI; H: HindIII; B: BamHI; nos: nopaline synthase; ApR: gen khng ampicillin;KmR: gen khng kanamycin

Mt v d chi tit v vector cis l pGV3850 (Zambryski, 1983). Vector ny cloi b cc gen onc ca Ti-plasmid kiu nopaline (C58) v thay bng pBR322 (Hnh7). Bt k trnh t DNA ngoi lai no c to dng trong pBR322 u c tha vo pGV3850 bng qu trnh chuyn vo Agrobacterium theo cch tip hp ti thp bao gm hai bc. Trc ht pBR322 mang gen cn chuyn v mt marker khngthuc khng sinh cho php chn lc Agrobacterium (kanamycin hoc streptomicin/spectinomycin) c a vo dng Ecoli cha hai plasmid h tr pGJ28 v pRGdrd11.Cc plasmid ny cung cp ColE1 c chc nng h tr, cho php chuyn c 3 plasmid voAgrobacterium qua tip hp. Tuy nhin pBR322 khng th ti bn trong Agrobacteriumv v vy n s khng c bo tn, nhng s ti t hp c th xy ra gia pBR322 vpGV3850 lm cho gen quan tm c chuyn vo Ti-plasmid. Th nhn tip hp c thc chn lc nh tnh khng do plasmid m ha v tnh khng rifampicin do nhim scth ca Agrobacterium m ha.

Vector nh th

Vector trans hoc vector nh th c da trn cc plasmid c th ti bn c E.coli vAgrobacterium v cc plasmid ny c cha cc trnh t bin ca T-DNA (Hnh 6B). Ccvector ny c th c thit k sao cho cc trnh t bin cnh MCS cho php xen DNAngoi lai vo v cc marker cho php chn lc trc tip cc t bo thc vt c bin


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np. Plasmid c th c thao tc trong E.coli v c chuyn vo qua s tip hpvi cc dng Agrobacterium mang plasmid c cha vng vir nhng thiu T-DNA v cctrnh t lp 25 bp. Cc plasmid nh th thng l cc th t bin mt on n ginca Ti-plasmid octopine kiu di hoc kiu nopaline . Vic chuyn DNA ngoi lai trnvector to dng vo t bo thc vt c th c thc hin do hot ng chc nng cavng vir.

pBin19 l mt vector nh th ph bin da trn n v ti bn (replicon) vt ch m rngca pRK252 . V vy n c th ti bn trong c E.coli v Agrobacterium, cho php xenDNA ngoi lai vo vector v sng lc trong E.coli trc khi chuyn vo Agrobacterium.Vector ny cha marker khng kanamicin (K mr)

Cu trc v s s dng vector nh th pBin19

A. Bn ca vector nh th pBin19

LB: bin tri; RB: bin phi; lac: vng b sung ca operon lac; Pnos: promoter gennopaline synthetase; npt-II: trnh t m ha neomycin phosphotransferase-II t Tn5;nosA: trnh t polyadenyl ha gen nopaline synthase.


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B. S xm nhp ca pBin19 vo A.tumefaciens LBA4404 s dng plasmid h trpRK2013 v sau loi b plasmid h tr trong Agrobacterium.

gip chn lc trc tip trong vi khun cng nh cc trnh t bin T-DNA cnh mtmarker chn lc tri vi mc ch s dng trong cc t bo thc vt (mt gen khngkanamicin nm cnh promoter nos v trnh t poly(A) thm vo) v mt MCS trongvng b sung ca -galactosidase. S sng lc cc th ti t hp c th c tinhnh Lac- E.coli nh s b tr vi s c mt ca IPTG v X-GAL (Gronebornv Messing, 1978). Vector ny c th c a vo Agrobacterium bng s tip hps dng cc chc nng h tr ca pKR2013. Agrobacterium ch LBA4404 cha Ti-plasmid (pLBA4404) loi b T-DNA nhng vng vir vn khng i. Th nhn tiphp c th c chn lc nh tnh khng kanamicin v rifampicin.

H thng vector nh th c trnh by bng trn. Cc vector nh th ny khc nhau vkch thc, s ti bn n nh trong Agrobacterium, cc v tr to dng, s b tr c th sng lc plasmid ti t hp trn cc a X-GAL, cc gen marker chn lc ccthc vt bin np v cc gen marker chn lc cc th nhn tip hp, nhng phn lnc bit l thch hp vi mt s plasmid h tr mang gen vir ca A.tumefacien cngnh vi nhiu Ri-plasmid.

Vector bin np thc vt s dng A.rhizogenes

Cc vector ny c s dng ln u tin khi bin np vo cc loi thc vt m s tisinh ton b c th t cc t bo n l l kh khn. mt s loi, s ti sinh c th cth xy ra t s nui cy r gy ra bi A.rhizogenes thng qua s pht trin phi somahoc s pht sinh c quan. C ch kiu hnh lng r b c ch lm pht sinh hnh thichi hin nay cha r. Thc vt ti sinh t lng r thng b bin i hnh thi: l b gpnp, h thng r n nghing, sinh trng cn ci v kh nng sinh sn km.

Vector lin hp

y l vector trung gian c thit k cho php thao tc E.coli v cha vng T-DNA ca Ri-plasmid. S dn nhp vector vo A.rhizogenes mang mt Ri-plasmid bnhthng lm n nh cc trnh t ca vector trung gian l kt qu ca ti t hp tngng ni phn t trong Ri-plasmid (Jensen, 1986).

Vector nh th

Vector nh th loi b ht kh nng gy hi c ngun gc t A.tumefaciens (Bevan,1984; Van den Elzen, 1985) c th a vo A.rhizogenes v s ti bn n nh min ls chn lc c duy tr. Sau nhim A.rhizogenes mang vector nh th vi thc vts lm chuyn T-DNA t vector nh th cng vi T-DNA ca A.rhizogenes vo genomethc vt (Shalin, 1986; Simpson, 1986). Cc gen ny khi chuyn vo genome thc vt


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c nh v gia cc trnh t bin ca vector nh th. Cc trnh t bin ca vector nhth ny c tc dng sau khi s cm ng ca vng vir ca Ri-plasmid kiu di ngay tich (Simpson, 1986; Trulson, 1986). iu ny c th xy ra l do tng ng ln, mc DNA, gia cc vng gy c (virulence region) ca A.tumefacien v A.rhizogenes(Huffman, 1984).

Gn y lng t (hairy root) c chua v da chut c to ra bi A.rizhogenes kiudi mang vector nh th vi marker chn lc Kmr. Th ti sinh t mi trng nui cylng r khng kanamicin ny c sng lc i vi thc vt kiu hnh bnh thngv mt s c phc hi m thiu T-DNA ca A.rizhogenes nhng c mt T-DNAca vector nh th (Shalin, 1986; Trulson, 1986) l do s chuyn c lp ca cc T-DNAtch ri. c tnh ny ca A.rhizogenes lm cho n tr nn hp dn nh l mt vectorbin np nu nh qu trnh ny c th c m rng i vi nhiu loi cy trng khc.


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Cc vector s dng chuyn gen thc vtCc vector bin np vo t bo thc vt s dng AgrobacteriumS gy ra cc khi u do AgrobacteriumTi-plasmid ca Agrobacterium tumefaciensRi-plasmid ca Agrobacterium rhizogenes

C ch chuyn T-DNAPlasmid Agrobacterium l vector bin npCc thnh phn c bn ca vector Ti-plasmid khng gy ung thCc vector bin np thc vt khng gy ung th da trn Ti-plasmidCis vectorVector nh th

Vector bin np thc vt s dng A.rhizogenesVector lin hpVector nh th

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