CNG NGH GEN (SV T TM TT LM)

Cng ngh gene: l ng dng cc k thut v sinh hc: gene, protein ci bin di truyn ca 1 sinh vt no . To ra sinh vt c gene bi bin i phc v nhu cu ca con ngi. S KHI QUT k thut gene(kt ti t hp DNA c thc hin qua nhiu cng on phc tp v tinh vi, c th ni l mt cng ngh vs cc bc ch yu sau: Bc 1: nui t bo ch to vector chuyn gene v nui t bo cho(nh tb ngi) cung cp DNA Bc 2: tch DNA plasmid v DNA t bo cho (ngi) Bc 3: ct c 2 loi DNA bng cng 1 loi enzyme restriction endonuclesae EcoRI to cc u so le c kt. Bc 4: trn chung DNA plasmid b ct on vs DNA ca t bo cho chng gn vo nhau v bt cp b sung nh cc u so le c kt. Bc 5: thm enzyme ni ligase to lin kt ha tr thnh DNA ti t hp hon chnh. Bc 6: bin np DNA ti t hp vo tb nhn Bc 7: chn lc v to dng vi khun mang gen l. sau khi c dng s to iu kin gen biu hin to ra sn phm.

Cc lnh vc chnh ca cng ngh gen: 1. To vt liu di truyn: DNA ti t hp ADN ti t hp l phn t DNA c to thnh t hai hay nhiu trnh t ADN ca cc loi sinh vt khc nhau. K thut DNA ti t hp c to ra ct dn mt phn on DNA l vo phn t nh c kh nng sao chp (nh plasmid vi khun), sau s khuch i phn on ny cng vs n v cho kt qu mt dng phn t phn on chn DNA.

Quy trnh: Tch DNA NST ca t bo cho v tch plasmid ra khi t bo vi khun. Ct v ni ADN ca t bo cho vo ADN plasmit nhng im xc nh, to nn ADN ti t hp. Thao tc ct tch on ADN c thc hin nh enzim ct (restrictaza). Cc phn t enzim ny nhn ra v ct t ADN nhng nucltit xc nh nh ngi ta c th tch cc gen m ho nhng prtin nht nh. Vic ct t ADN vng ca plasmit cng c thc hin do enzim ct cn vic ghp on ADN ca t bo cho vo ADN plasmit th do enzim ni (ligaza) m nhim. Chuyn ADN ti t hp vo t bo nhn, to iu kin cho gen ghp c biu hin. Plasmit mang ADN ti t hp c chuyn vo t bo nhn bng nhiu phng php khc nhau. Vo t bo nhn, n t nhn i, c truyn qua cc th h t bo sau qua c ch phn bo v tng hp loi prtin m ho trong on ADN c ghp. T bo nhn c dng ph bin l vi khun ng rut E.Coli. T bo E.Coli sau 30 pht li t nhn i. Sau 12 gi, 1 t bo ban u s sinh ra 16 triu t bo, qua cc plasmit trong chng cng c nhn ln rt nhanh v sn xut ra mt lng ln cc cht tng ng vi cc gen ghp vo plasmit. Trong k thut cy gen ngi ta cn dng th thc khun lm th truyn. N gn on ADN ca t bo cho vo ADN ca n v trong khi xm nhp vo t bo nhn n s em theo c on ADN ny vo .

2. To ging SV chuyn gen: To ging ng vt chuyn gen, dng sinh vt bin i gen, ging cy trng bin i gen. S dng cng ngh gen to ra nhng ging V mi c nng sut v cht lng cao hn v sn phm, c bit to ra ng vt chuyn gen c th sn xut ra thuc cha bnh. a, To ging cy trng bin i gen: Chuyn gen vo thc vt l mt trong nhng lnh vc pht trin nhanh nht ca cnsh phn t. T nm 1980, nh vt qua kh khn k thut, k thut gen c nhng bc tin ngon mc v t n th h th 3: -bc 1: ci thin ging cy trng: a vo cc ging cy trng cc n gen nh khng thuc dit c v khng bnh. Gn y c cc n lc hng vo s chuyn a gen.gm: to cy trng c kiu hnh khng virus, to ging khng nm, chuyn gen khng su hi v d chuyn gen Bt ca Bacillus thuringiensis vo c chua, thuc l, bng.to sc t thc vt chuyn gen - bc 2: bin i cht lng thc phm ca cy trng: bin i hng lot tnh cht nh tng amino acid c gi tr, lipid tt hn, tng lng vitamin, ngt, cht bt.

V d: ging la c chuyn gen tng hp caroten, sau qu trnh tiu ha, carotene c chuyn ha thnh vitamin A. Cng ngh to ra la vng da trn nguyn tc la tch t carotene trn l, khng phi trong ht go. Nh thm vo hai gen phytoene synthase v phytoene desaturase bng phng php chuyn gen, carotene ny s chuyn v c vo trong phi nh ht go. - bc 3: thc vt sn xut cc loi ha cht c bit Du nhn cng nghip:cc ging ci du chuyn gen c lipid bin i trong ht Cht do plastic: cy Arabidopsis mang gen vi khun sn xut cht do PHB. Nm 1996, c thng bo nhn c 1 kg cht do PHB t cy thuc l chuyn gen. Cc protein tr liu: to thc vt sn xut enkephalin, serum albumin, -xu hng hin nay:l ch trng chuyn gen ngi vo thc vt do nhiu li im: c th trng vs quy m ln nh nng lng mt tri, t tn km hn, virus thc vt ko ng s cho ngi,cho gen biu hin m to du thc vt nn rt d tch v thu nhn protein . b, To ging ng vt chuyn gen Phng php to ng vt bin i gen: - y trng ra khi con vt no -Cho th tinh trong ng nghim. -Tim gen cn chuyn vo hp t v hp t pht trin thnh phi. -Cy phi c chuyn gen vo t cung con vt khc mang thai v sinh bnh thng. Sinh ra mt con vt chuyn gen

Cc ng dng: Cc ng vt mang gen ngi lm m hnh th nghim: hng lot chut mang bnh ung th, Alzheimer, thoi ha c, vim khp,... Cc chut chuyn gen cng c dng lm cc h thng th nghim nh cc dng mang gen m ha CFTR protein ngin cu tr bnh ha x nang CF Cc ng vt nui chuyn gen sn xut r-protein: cc v chuyn gen (b, d, cu, heo, th) chuyn gen sx r-protein cho ngi nh cho gen biu hin tuyn sa chn nui gen sn xut vi chc loi protein tr liu.

3. To khuynh hng cha tr s dng cng ngh gene. Cc xt nghim SNP: vic gii trnh t b gen ngi m ra hng ngn thng tin v cc SNP hu ch. Cc xt nghim SNP gi vai tr then cht trong vic thit k thuc cho tng c nhn. Dc hc b gen: - Pharmacogenomics: l s kt hp gia dc hc vs genomics nhm nghin cu v gen, s iu ha v ng gp ca chng vs s pht trin cu bnh tt mi c th ring bit. - mt lnh vc mi khc l toxicogenomics. N xc nh s nhy cm c th vs cc cht nhim mt or thuc. Cng ngh gen tc ng n tng ngi: ngay khi d n b gen ngi bt u, xut hin cc cm t tr em theo n t hng hay thit k tr emm ngay trong th k 21 ny c th thnh hin thc. VIT NAM pht trin nht l cng ngh to ging cy trng chuyn gen. - vic ngin cu ang dc tin hnh cc phng th nghim, ng thi m rng quan h hp tc vs cc t chc nghin cu quc t v cc cng ty nc ngoi nn ko trnh khi du nhp cy trng v cc sp chuyn gen. - trc mt, nc ta tp trung chuyn gen c tnh khng cn trng, khng bnh virus vo nhng i tng cy trng ko lm thc phm gm loi cy cng nghip nh: bng vi, hoa cnh, cy lm nghip nh: bch n, thng, hng, keo, bc u th nghim cc cng ngh tin tin nh iRNA vo vic to tnh khng bnh virus. Mt s PTN cng ngin cu chuyn gen khng nguyn ca virus vo cy trng, th nghim phng thc sx vaccine dng ng ming. V D: SN XUT INSULIN TI T HP Insulin (C257H383N65O77S6, M=5808) l mt loi hormone do cc t bo o ty ca tuyn ty tit ra vs tc dng chuyn ha carbohydrate. Ngoi ra, insulin cn c tc dng n chuyn ha m m v gan thnh nng lng ATP cung cp cho cc hot ng sng ca c th. u tin insulin c tch chit t tuyn ty ca ng vt, y l gii php u tin cho bnh tiu ng type I. Thi k tip theo l s t ph trong k thut DNA ti t hp vs thnh cng u tin l insulin tng hp ca ngi c sn xut.

Nm 1978, on gen m ha cho phn t insulin ngi c cc nh khoa hc thuc cng ty Genetech tch dng v chuyn vo vk E.coli. on gen ny dc ghp vo plasmid of E.coli plasmid ti t hp mang gen insulin plasmid bin np vo vk to dng vk sn sinh insulin.

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