Qhov Tseem Ceeb Qhov txawv - Gene Cloning vs PCR
Qhov kev sib txuas ntawm ntau cov ntawv luam ntawm DNA los ntawm ib qho DNA tshwj xeeb yog hu ua DNA amplification. Muaj ob lub ntsiab DNA amplification txheej txheem uas yog noob cloning thiab PCR. Qhov sib txawv tseem ceeb ntawm gene cloning thiab PCR yog, gene cloning tsim ntau cov ntawv luam ntawm ib lub noob tshwj xeeb hauv vivo los ntawm kev tsim cov DNA recombinant thiab loj hlob hauv cov kab mob hauv tsev thaum PCR tsim ntau lab daim ntawv luam ntawm ib qho DNA tshwj xeeb hauv vitro rov ua dua ib zaug. denaturation thiab synthesis.
Gene Cloning yog dab tsi?
Gene cloning yog cov txheej txheem ua haujlwm los nrhiav thiab muab cov noob tshwj xeeb los ntawm kev rho tawm genomic DNA ntawm cov kab mob los ntawm kev tsim cov DNA recombinant. Genomic DNA muaj ntau txhiab tus noob sib txawv uas tau muab tso rau hauv cov proteins. Thaum DNA tau muab rho tawm, nws suav nrog tag nrho cov noob caj noob ces nws tuaj yeem dais. Gene cloning txheej txheem tau ua kom pom cov noob tshwj xeeb los ntawm tag nrho DNA. Yog li ntawd gene cloning yog ib qho cuab yeej tseem ceeb hauv molecular biology.
Kev ua lub tsev qiv ntawv genomic ntawm ib yam kabmob yog qhov tseem ceeb hauv cov noob qoob loo yog tias tsis muaj qhov tsis paub txog qhov chaw ntawm cov noob cuam tshuam hauv DNA. Lub tsev qiv ntawv genomic yog tsim los siv cov kauj ruam hauv qab no.
Kauj Ruam 1: Kev rho tawm tag nrho DNA los ntawm cov kab mob uas muaj cov noob xav tau.
Kauj Ruam 2: Kev txwv kev zom cov DNA rho tawm los tsim cov khoom me me. Cov kauj ruam no yog pab txhawb los ntawm kev txwv endonucleases.
Kauj Ruam 3: Xaiv cov vector tsim nyog thiab qhib vector DNA siv tib yam kev txwv endonucleases. Cov kab mob plasmids feem ntau yog siv los ua cov vectors nqa cov DNA txawv teb chaws. Plasmids yog lub voj voog me me ntawm DNA nyob hauv cov kab mob.
Kauj Ruam 4: Kev sib xyaw ntawm vector DNA thiab tawg DNA los tsim cov DNA molecule recombinant. Cov kauj ruam no yog tswj hwm los ntawm DNA ligase.
Kauj Ruam 5: Hloov pauv ntawm cov DNA molecules recombinant rau hauv cov kab mob hauv tsev. Cov kauj ruam no yog hu ua kev hloov pauv, thiab ua tiav los ntawm kev siv hluav taws xob poob siab.
Kauj Ruam 5: Kev tshuaj ntsuam ntawm cov kab mob hloov pauv ntawm cov kab lis kev cai nruab nrab. Cov neeg sib xyaw ntawm cov hloov pauv thiab tsis hloov pauv cov tswv tsev yog tau txais thaum kawg ntawm txoj kev hloov pauv. Raws li gene ntawm kev txaus siab muaj xws li tsuas yog nyob rau hauv hloov pauv host cells. Yog li ntawd, nws yog ib qho tsim nyog los xaiv cov hloov pauv hloov pauv. Kev xaiv yog tsim los ntawm kev xaiv cov xov xwm uas muaj cov tshuaj tua kab mob. Tsuas yog cov hlwb hloov pauv loj hlob ntawm qhov kev tshuaj ntsuam nruab nrab no ua rau kev xaiv.
Kauj Ruam 6: Kev loj hlob ntawm cov kab mob los tsim cov tsev qiv ntawv gene. Hauv cov kauj ruam no, cov hloov pauv cov tswv tsev tau nkag mus rau hauv cov kab lis kev cai tshiab uas muab kev loj hlob zoo tshaj plaws. Tag nrho cov cheeb tsam ntawm daim phiaj kab lis kev cai sawv cev rau lub tsev qiv ntawv genomic ntawm lub cev.
Kauj Ruam 7: Cov DNA molecule recombinant uas muaj cov noob ntawm kev txaus siab yuav tsum tau tshuaj xyuas los ntawm ntau txhiab cloned fragments ntawm recombinant DNA. Nws tuaj yeem ua tiav los ntawm kev siv cov tshuaj ntsuam xyuas uas cim cov noob tshwj xeeb lossis cov protein tshwj xeeb los ntawm cov noob ntawd.
Thaum tus neeg nyiam cov noob muaj cov kab mob colony raug txheeb xyuas los ntawm tag nrho cov cheeb tsam, nws muaj peev xwm ua tau ntau lab luam ntawm cov plasmid recombinant uas muaj cov noob.
Gene cloning yog siv los tsim cov tsev qiv ntawv gene, tsim cov protein tshwj xeeb, vitamins, tshuaj tua kab mob, tshuaj hormones, sequencing thiab mapping genomes ntawm cov kab mob, ua ntau cov ntawv luam ntawm tib neeg DNA hauv forensics thiab lwm yam.
Figure_1: Gene Cloning
YPCR yog dab tsi?
Polymerase Chain Reaction (PCR) yog cov txheej txheem uas tsim ntau cov ntawv luam ntawm ib qho DNA tshwj xeeb. Exponential amplification ntawm ib qho DNA ib ntus yog tau los ntawm PCR nyob rau hauv vitro tej yam kev mob. Cov txheej txheem no yog ib qho cuab yeej muaj zog heev hauv Molecular Biology vim nws tuaj yeem muab cov qauv me me ntawm DNA rau hauv qhov siv tau. PCR tau qhia los ntawm Kary Mullis hauv xyoo 1983 thiab qhov khoom plig tau txais txiaj ntsig no tau tsim kev nce qib loj hauv Molecular Biology.
PCR cov txheej txheem ua raws li kev rov ua dua PCR cov tshuaj tiv thaiv raws li qhia hauv daim duab 02. Ib qho tshuaj tiv thaiv PCR muaj peb theem tseem ceeb tshwm sim ntawm peb qhov kub sib txawv; denaturing ntawm ob stranded ntawm DNA ntawm 94 0 C, annealing ntawm primers ntawm 68 0 C thiab strand elongation ntawm 72 0 C. Yog li ntawd, thaum PCR ua tiav, qhov kub thiab txias hloov pauv yuav tsum tau tswj xyuas kom zoo dua. PCR yog ua nyob rau hauv lub tshuab PCR hauv PCR hlab. PCR tubes yog loaded nrog PCR cov khoom sib xyaw uas muaj cov qauv DNA, Taq polymerase, primers, dNTPs thiab tsis. Denaturing ntawm ob chav stranded qauv DNA rau hauv ib leeg stranded DNA yog ua los ntawm kev rhuav cov hydrogen bonds ntawm complementary bases ntawm 94 - 98 0 C. Tom qab ntawd ib leeg ntawm cov qauv DNA raug nthuav tawm rau cov primers. Ib nkawm primers (rau pem hauv ntej thiab thim rov qab) yuav tsum tau muab, thiab lawv yuav tsum tau thermostable kom zam tau qhov kub thiab txias. Primers yog ib qho stranded luv DNA sequences ntxiv rau qhov kawg ntawm lub hom phiaj DNA tawg. Synthetic primers yog siv hauv PCR. Primers khi nrog lub hauv paus ntxiv ntawm cov qauv DNA thiab pib qhov kev sib txuas ntawm cov hlua tshiab. Cov kauj ruam no yog catalyzed los ntawm ib qho enzyme hu ua Taq polymerase; Ib qho thermostable DNA polymerase enzyme cais los ntawm Thermus auqaticus. Thaum primers thiab nucleotides (cov tsev thaiv) muaj, Taq polymerase tsim cov strand tshiab ntawm DNA ntxiv rau cov qauv DNA. Thaum kawg ntawm PCR program, amplified DNA fragment yog pom siv gel electrophoresis. Yog xav tau kev tshuaj xyuas ntxiv, cov khoom PCR raug ntxuav los ntawm cov gel.
PCR yog qhov muaj txiaj ntsig zoo rau kev kuaj mob thiab saib xyuas cov kab mob caj ces thiab kis tau tus kab mob, txheeb xyuas cov neeg ua txhaum cai (hauv thaj chaw forensics), kawm txog cov qauv thiab kev ua haujlwm ntawm lub hom phiaj ntawm DNA, kev sib txuas thiab kev kos duab ntawm genomes ntawm cov kab mob, thiab lwm yam. PCR tau dhau los ua cov txheej txheem kuaj niaj hnub hauv kev tshawb fawb txog kev kho mob thiab molecular biology kuaj ntawm cov kws tshawb fawb vim nws muaj ntau yam kev siv.
Figure_2: Polymerase Chain Reaction
Qhov txawv ntawm Gene Cloning thiab PCR yog dab tsi?
Gene Cloning vs PCR |
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| Gene cloning yog txheej txheem ntawm kev ua ntau daim ntawv theej ntawm cov noob tshwj xeeb hauv vivo los ntawm cov DNA recombinant thiab hloov mus ua tus kab mob. | Cov txheej txheem PCR tsim ntau daim ntawv theej ntawm DNA ib ntus hauv vitro los ntawm kev rov ua dua ntawm PCR cov tshuaj tiv thaiv. |
| Yuav tsum tau tsim kho DNA | |
| Recombinant DNA yog tsim los txhawm rau nrhiav cov noob. | Recombinant DNA tsis tsim. |
| Nyob hauv kev ua haujlwm | |
| txheej txheem no yog kev siv zog ntau. | tsis xav tau kev ua haujlwm hnyav. |
| In vivo lossis In vitro txheej txheem | |
| Kev tsim cov DNA recombinant yog nyob rau hauv vitro thiab lub amplification ntawm DNA yog nyob rau hauv vivo. | Kev nthuav dav ntawm DNA tshwm sim hauv vitro. |
Summary – Gene Cloning vs PCR
Gene cloning thiab PCR yog ob txoj hauv kev siv rau DNA amplification. PCR yog cov txheej txheem hauv vitro uas ua rau ntau daim ntawv theej ntawm DNA ntawm ib qho DNA tshwj xeeb yam tsis siv DNA rov ua dua thiab cov kab mob hauv lub cev. Gene cloning feem ntau yog cov txheej txheem hauv vivo uas ua rau ntau daim ntawv luam ntawm cov noob txaus siab hauv cov kab mob hauv lub cev los ntawm kev tsim kho ntawm DNA recombinant. Qhov no yog qhov txawv ntawm Gene cloning thiab PCR.
