NUI CY M T BO THC VT BNG T BO TRN V K THUT CHUYN GEN KHI NIM Nui cy m- t bo thc vt l k thut cho php nui

i cy d dng nhng t bo thc vt hay m phn sinh sch bnh trong mi trng nhn to thch hp to ra nhng khi t bo hay nhng cy hon chnh trong ng nghim. C S SINH L CA CNG NGH NUI CY M- T BO TNH TON NNG: L kh nng ca 1 t bo hnh thnh 1 cy ha chnh trong iu kin nui cy thch hp, do trong t bo c cha b AND(NST) hon chnh, cha ton b thng tin di truyn cho 1 chu k sng hon chnh. AND mARN Protin tnh trng Ty tng t bo, tng loi m, tng thi k sinh trng, pht trin m cc gen ph hp hot ng; cc gen khng cng hot ng nh nhau trong cc giai on pht trin ca c th (do c ch iu ha hot ng ca gen). S PHN CHIA, PHN HA, PHN PHN HA CA T BO: Phn ha: 1 t bo 1khi t bo(phn ha) m c quan h c quan. Phn phn ha: khi cc t bo phn ha thnh cc m chc nng ring bit nhng vn c th quay v trng thi chc nng phi sinh ban u ki gp iu kin thun li. Cc c tnh trn c xc nh da trn c s l c im di truyn nhng s thay i mi trng cng c nh hng trc tip n cc c tnh . CC NHN T M B THNH CNG TRONG CNG NGH NUI CY M- T BO THC VT: m bo iu kin v trng, phng th nghim phi chuyn ha cao. Chn ng mi trng v chun b mi trng ng cch. Chn m cy, x l m cy thch hp trc v sau khi cy.

CC BC TRONG K THY NUI CY M- T BO THC VT To vt liu khi u: thng chn chi l b phn nui cy thch hp nht kh trng cy trong mi trng khi ng ti sinh Giai on nhn nhanh: vt liu khi u c chuyn sang mi trng nhn nhanh c b sung Xitokinin ti sinh 1 thnh nhiu chi. Giai on to cy hon chnh: tch ring cc chi cho vo mi trng to r (b sung Auxin) mi chi ra r thnh 1 cy hon chnh. Giai on ra cy: cy trong ng nghim tiu chun (chiu cao, s l, s r) s c chuyn sang mi trng t nhin. CC HNH THC NUI CY M Nui cy m thc vt Nui cy m bng t bo trn NUI CY M THC VT Nui cy m ca c quan tch ri: VD: c c rt ct lt tch mnh mch ry x l+ nui cy m phi phi ny mm cy non trong ng nghim cy trng thnh Nui cy t m hay c quan: VD: t 1 m l tch t bo nui cy trn a Ptri iu kin thun li s hnh thnh nhng cy non trng chu pht trin cy trng thnh. Nui cy nh sinh trng: T 1 m phn sinh ca nh sinh trng ca r hoc thn nui cy trong mi trng dinh

dng s to nn cy con Nui cy m so t ht phn( Auxin rt quan trng): Nui cy bao phn c ht phn chn mi trng thun li bao phn chn s phn ha bng cch: nhn sinh sn s tiu bin nhn sinh dng s nguyn phn to m so nui dng cy n bi(n) x l cnxixin cy trng thnh(n bi kp). NUI CY M BNG T BO TRN Khi nim t bo trn: L t bo thc vt b loi b vch bi 1 x l enzim. l t bo t do, c lp, khng nh hng( v khng cn chu s tng quan trong h thng thc vt). S C LP V NUI CY T BO TRN S c lp: Trc ht phi x l vt liu khi u( l, m so, dch treo t bo) vi enzim phn hy vch( Xenlulaza, HemiXenlulaza, Pectolyaza). V d: phng php to t bo trn t l: L lm sch l bng Clorox 10% ra li l 3 ln bng nc ct tit trng tch lp biu b di ca l+ gy co nguyn sinh 1h bng Mannitol 10% enzim thu c t bo trn. Sc trng t bo bnh thng th c cn bng vi sc p c hc ca vch vch mt, t bo khng b v th cho vo nhng cht gy co nguyn sinh (CaCl2,KCl,Mannitol) ha tan trong Enzim khi x l S NUI CY Thnh phn ca mi trng nui cy lng hay c ty thuc vo vat liu thc vt.Mi trng ny c thm Auxin v Xitokinin gip s ti to vch v cc ln phn cia u tin m bo cc yu t dinh dng: axit amin, polyamin, Hydrolysat Cazein, nc da, mch nha m bo iu kin nhit , PH, nh sng,p sut thm thu. Trong ln t nhn i u tin, mi trng phi c p sut thm thu cao, Auxin, Xitikinin thch hp, nh sng yu. Sau cn gim p sut thm thu bng cch pha long mi trng gip cho s tng trng t bo. Khi m so c hnh thnh cn chuyn chng vo trong mi trng rn cha Auxin nng thp hn v Xitokinin cao hn. Sau cng kch thch ra r cn loi Xitokinin, tng nng Auxin. Thng thng, t bo trn cn 1-2 ngy cho s hnh thnh vch, 2-7 ngy cho cc ln nhn i u tin, 3-4 tun cho s tng trng m so, sau 1-2 thng c phi hnh cu, 1thng na c cy ny mm. CH LI CA T BO TRN T bo trn c kh nng ti to vch v cho phi th h. Nui cy t bo trn s m ra 1 m hnh hp dn theo di qu trnh sinh phi th h t 1 t bo c lp. c bit bit c s sp xp cc si Xenluloz xc nh hng ko di t bo, v tr v hng ca mt phng phn chia. Thun li cho s nghin cu v sinh l t bo: tnh thm ca mng, vn chuyn cc cht ha tan, ion, c ch hot ng ca hoocmon thc vt T bo trn c th chu cc x l gy t bin v c dng to ra cc dng t bo c ngun gc n bo. T bo trn thit yu cho s lai th h( dung hp t bo trn) v p dng cc k thut chuyn

gen( bm AND, ha thm, in thm) M RNG NUI CY M BNG K THUT DUNG HP T BO TRN V K THUT CHUYN GEN S LAI TH H BNG K THUT DUNG HP T BO TRN Khi nim: Dung hp l hin tng ct t mng sinh cht ni tip xc gia 2 t bo trn khc loi do tc ng ca cc nhn t bn ngoi. Sau l s ti t chc cc mng ban u thnh 1 v bao ly t bo cht v 2 nhn cha m. CC HNH THC DUNG HP T dung hp: V mng sinh cht ca t bo(-) do nhm P v Protein nn cc t bo trn y nhau nn khng t dung hp. Do phi lm cc r bo trn tip xc nhau nh PEG hay sc in. Ha dung hp: S ha dung hp c th thc hin vi NaNO3 0,25M, PVA 15 %nhng tn s dung hp thp nn t c s dng. Virut Senday gim hot tnh l cht thng dng hin nay bng cch cho git hn hp 2 loi t bo trn tip xc t t vi cc git PEG 25- 50% in dung hp (ttnht): Khi t hn hp 2 loi t bo trn trong 1 in trng, cc t bo trn s di chuyn, tip xc nhau v xp thnh 1 chui di di lc in trng Sau dng xung in ngn to vt t mng, s dung hp s xy ra ti ni tip xc. NHNG BIN I CA T BO Phn ln t bo lai c b NST khng bn, nht l t cha m xa nhau, 1 vi NST b hy b sau 1 vi ln phn bo. Tnh bn ch thy c sau nhiu ln nhn i. Dung hp t bo trn c th theo 2 kiu: Dung hp nhn 1 phn hay ton phn: hnh thnh t bo lai. 1 trong 2 nhn cha m c gi: t bo cht lai C 2 u mang gen trong t bo cht. CH LI V TR NGI CA NUI CY T BO TRN ch li: gip loi tr tnh bt th hu tnh, to cy lai hu th; Gip chuyn nhng c tnh c li vo cy trng, t i hi phng tin phc tp; Khng cn hiu bit chnh xc v s lin h gia cc gen, t tn km, nhanh v trc tip. Tr ngi: Qu trnh c lp nui cy phi hon thin. Cha c phng php hiu qu tuyn chn cc sn phm ph hp. S CHUYN GEN TRONG K THUT NUI CY M CC KHI NIM V K THUT DI TRUYN Lai phn t: L s dung hp hai phn t ADN c ngun gc khc nhau hnh thnh mt phn t ADN lai( ADN ti t hp) hoc hnh thnh mt phn t ADN lai gm hai mch: mt mch l chui PolyNu, mch i xng l chui PolyRinu Cong nghe di truyen (ng dung cua k thuat di truyen) : La quy trnh s dung k thuat di truyen e san xuat san pham sinh hoc tren quy mo cong ngiep e ap ng nhu cau cho con ngi. La qua trnh co tnh toan en san pham, gia thanh, kinh te. K thut di truyn: L k thut thao tc trn vt liu di truyn da trn nhng hiu bit v cu trc ca AxitNucleic v di truyn vi sinh vt. S truyn Gen:

L s bin i b gen t bo bi s xen vo 1 on ADN ngoi lai sinh. ADN ti t hp: L phn t ADN lai t c do s kt hp gia 2 trnh t ADN ca 2 loi khc nhau, hoc cng c th c hnh thnh t 1 mch ARN v 1 mch AND. Cc Enzym cng c: Enzym ct(restriction Enzym): c kh nng nhn bit v ct ADNsi kp bhng v tr rt chnh xc. Ligaza: ni lin 2 on ADN bng cch to cc mi lin kt P dieste lin kt cc Nu. Reverse transcrip tase: cho php sao chp ngc( tc s tng hp cADN t mARN. Vct truyn: Phn t ADN nh c dng ghp on ADN cn nghin cu gi l vct truyn. Thng dng l Plasmid vi khun hay ADN virut. S to dng gen: L qu trnh c lp v thu nhn nhiu bn sao ging nhau ca 1 gen hay 1 on gen da trn kh nng t nhn i ca ADN. y l c s cho s tng bi on ADN cn nghin cu. S sng lc v d phn t: S sng lc l qu trnh tuyn chn cc Vect cha on ADN cn nghin cu. S tuyn chn thc hin nh lai phn t kt hp vi 1 d phn t. D phn t: L cc on axit Nucltit( ADN, ARN) si n: 20 n vi trm Nu; Ngi ta dng nh du on axit Nucltit cn nghin cu trong 1 phn ng gi l lai phn t nh c nh du phng x v c tnh b sung v i song vi 1 vng c on A.Nu cn nhn bit. S CHUYN GEN GIN TIP BI AGROBACTERIUM S CHUYN GEN TRC TIP(BIN NP) Cc phng php chuyn gen trc tip: ho thm, in thm, sng bn ADN: Ho thm: dng PEG v dng t bo trn.(tn s chuyn gen thp) in thm: dng xung in cao p v t bo trn. C s c in thm l khi t bo chu s tc ng ca xung in cao p, cc l nh tm thi c to thnh trn mng tr nn thm, ADN vo t bo c theo khuynh hng nng . Tuy nhin s l c to thnh tng theo in th cho ti 1 gi tr ti hn( qu im ny mng s b tn thng khng hon nghch). S thnh lp cc l nh c th do: in trng lm thay i cn bng in tch lp P lipit mng Tnh lng to nn nhng ch yu tm thi d b t di nh hng ca in trng. K THUT BN ADN Cc Plasmid c mang cc gen l c c inh trn cc ht kim loi( vng hay tungsten) c ng knh 0.000006m nh lc ht tnh in. Cc ht kim loi- Plasmid ny c bn vi vn tc ln xuyn qua vch, mng v xen vo b gen nhn t bo ch. Mng t bo c 1 tnh do hon ho nn d dng sa cha sau khi b tn thng. Cc phn t ADN tch khi ht kim loi sau khi vo nhn t bo gia nhp vo ADN NST ca t bo. u im: Nhanh, d thc hin, t cng on, hiu qu kinh t cao khng chuyn bit cho loi Khng i hi c lp v nui cy t bo trn. D pht hin cc t bo c mang gen c chuyn. Nhc im: Thit b t tin. Tn s chuyn gen thp.

C phng php gin tip v trc tip: sau khi t bo thc vt mang gen c tnh, chng s nguyn phn nhiu ln to ra1 khi t bo phn ho to m c quan cy trng thnh. Thanh tu ng dung cua k thuat nuoi cay mo bang te bao tran va k thuat chuyen gen Bang phng phap dung hp te bao tran va ap dung k thuat chuyen gen cac nha khoa hoc a vt qua moi rao can sinh hoc trong t nhien e tao cac te bao lai va cac giong thc vat mang cac gen mong muon. Gop phan quan trong trong viec cai tien giong cay trong, tang nang suat. Tng tnh chng chu vi cc iu kin kh hu, th nhng bt li Tng kh nng khng thuc dit c Tng kh nng chng chu vi cc cn trng v b cnh cng (bng cch chuyn gen m ho tng hp thuc tr su trong cy thc vt) Tng kh nng phng tr bnh cy Tng nng sut cy trng To thc n cha khng th c hiu cho gia sc Tuy nhin cng c mt s hn ch nh hng sc kho cng ng nh hng mi trng sinh thi A?ng kinh t chnh tr x hi