Video: Yuav ua li cas tom qab cov kab mob hloov pauv?
2024 Tus sau: Stanley Ellington | [email protected]. Kawg hloov kho: 2023-12-16 00:17
Kab mob tuaj yeem nqa DNA txawv teb chaws hauv cov txheej txheem hu ua kev hloov pauv . Kev hloov pauv yog ib kauj ruam tseem ceeb hauv DNA cloning. Nws tshwm sim tom qab txwv digest thiab ligation thiab hloov pauv tshiab ua plasmids rau kab mob . Tom qab kev hloov pauv , kab mob raug xaiv rau ntawm daim phiaj tshuaj tua kab mob.
Yog li, dab tsi yog qhov tseem ceeb ntawm lub sijhawm rov qab los tom qab kev hloov pauv?
Cell lub sijhawm rov qab los Tom qab kub poob los yog electroporation, hloov cov cell yog kab lis kev cai hauv cov kua tshuaj tua kab mob tsis muaj tshuaj tua kab mob rau lub sijhawm luv lub sij hawm kom tso cai qhia txog cov tshuaj tiv thaiv kab mob gene (s) los ntawm cov plasmid uas tau txais los pib (Daim duab 5). Cov kauj ruam no txhim kho cell viability thiab cloning efficiency.
Ib tug kuj yuav nug, puas yog kev hloov pauv kab mob ua haujlwm tau zoo? Tej yam cuam tshuam Transformation Efficiency Cov kev hloov pauv tshuaj tiv thaiv yog muaj txiaj ntsig thaum <10ng ntawm DNA siv. pUC19 DNA (0.1ng) yog tsim los tswj. Supercoiled DNA yog feem ntau muaj txiaj ntsig rau kev hloov pauv piv rau linear lossis ssDNA uas muaj cov transformation efficiency ntawm <1%.
Hais txog qhov no, vim li cas cov kab mob hloov pauv tseem ceeb?
Taw qhia. Kev hloov pauv yog tus txheej txheem uas txawv teb chaws DNA nkag mus rau hauv ib lub cell. Kev hloov pauv ntawm kab mob nrog plasmids yog tseem ceeb tsis yog rau kev kawm hauv kab mob tab sis kuj vim kab mob yog siv los ua ib txoj hauv kev khaws cia thiab rov ua cov plasmids.
Tus txheej txheem ntawm kev hloov pauv yog dab tsi?
Hauv molecular biology, kev hloov pauv yog qhov kev hloov pauv caj ces ntawm lub xov tooj ntawm tes uas tshwm sim los ntawm qhov ncaj qha uptake thiab koom ua ke ntawm exogenous genetic khoom los ntawm nws ib puag ncig los ntawm cell membrane(s).
Pom zoo:
Thaum ua qhov tshuaj ntsuam xiav dawb cov kab mob hauv cov kab mob muaj?
Hloov pauv, cov kab dawb dawb tsis tuaj yeem metabolize X-Gal los tsim cov xim xiav, vim tias lawv tsis tsim cov β-galactosidase ua haujlwm tom qab noj plasmid nqa cov DNA tso thiab cuam tshuam cov lacZ α noob. Cov kab mob dawb no muaj cov kab mob recombinant thiab yuav tsum tau xaiv (Daim duab 1)
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Cov kab mob hloov pauv li cas hauv chav kuaj?
Cov kab mob tuaj yeem nqa DNA txawv teb chaws hauv cov txheej txheem hu ua kev hloov pauv. Kev hloov pauv yog ib kauj ruam tseem ceeb hauv DNA cloning. Nws tshwm sim tom qab kev txwv kev zom thiab ligation thiab hloov cov plasmids tshiab rau cov kab mob. Tom qab kev hloov pauv, cov kab mob raug xaiv rau ntawm daim phiaj tshuaj tua kab mob
Dab tsi yog tus neeg sawv cev ntawm kev hloov pauv hauv cov kab mob?
Hauv cov kab mob, kev hloov pauv yog ua los ntawm kev sib xyaw transgenic DNA nrog cov kab mob hlwb kho kom lawv muaj peev xwm coj tau DNA. Ntau yam ntawm lwm txoj kev tau siv los qhia DNA rau hauv cov nroj tsuag thiab tsiaj txhu xws li DNA txhaj, electroporation, thiab microparticle bombardment