Qhov Tseem Ceeb - YAC vs M13 Phage Vector
DNA cloning yog txheej txheem tseem ceeb uas ua rau kev nthuav tawm ntawm DNA tseem ceeb ntawm cov kab mob. Nws yuav tsum tau koom nrog cov DNA tshwj xeeb nrog vector DNA los tsim cov DNA recombinant uas hloov mus rau hauv cov kab mob hauv lub cev. Ib tug vector yog DNA molecule uas coj zoo li lub tsheb nqa cov khoom siv txawv teb chaws mus rau lwm lub cell lossis kab mob. Nws yuav tsum muaj peev xwm rov ua dua hauv cov kab mob hauv lub cev thiab tsim ntau cov ntawv luam ntawm DNA recombinant. Muaj ntau hom vectors siv hauv DNA cloning. Poov xab dag chromosome (YAC) thiab M13 phage vector yog ob hom ntawm lawv. Qhov sib txawv tseem ceeb ntawm YAC thiab M13 phage vector yog tias YAC yog ib qho khoom siv chromosome uas rov ua dua nyob rau hauv cov poov xab thaum M13 phage vector yog ib txoj kab sib txuas DNA ntawm bacteriophage M13 uas replicates hauv E Coli hlwb.
YAC Vector yog dab tsi?
YAC yog ib qho khoom tsim los ntawm chromosome uas muaj peev xwm nqa tau ib ntu loj ntawm DNA txawv teb chaws thiab rov ua dua hauv cov poov xab. Nws muaj centromere, telomere thiab autonomously replicating sequences tseem ceeb rau replication thiab stability. YAC tseem yuav tsum dais cov cim xaiv lossis cov cim thiab cov chaw txwv kom ua rau nws ua tau zoo cloning vector. Cov kab ke loj xws li 1000 kb txog 2000 kb tuaj yeem muab tso rau hauv YAC thiab xa mus rau hauv cov poov xab.
Daim duab 01: YAC Vector
M13 Phaj Vector yog dab tsi?
Bacteriophage M13 yog tus kab mob uas kis tau thiab rov ua dua hauv E Coli. Lub genome ntawm M13 bacteriophage yog me me, txog 6.7 kb. Nws yog ib leeg-stranded, ncig thiab zoo siab DNA. Tus kab mob no tshwj xeeb kis kab mob E Coli los ntawm F pilus. Thaum nws nkag mus rau ssDNA rau hauv cov kab mob, nws ua ke nws cov hlua txuas ntxiv thiab dhau los ua dsDNA lossis daim ntawv rov ua dua (RF) ntawm M13. RF tuaj yeem coj tus cwj pwm zoo li plasmid hauv cov kab mob hauv lub cev. Lub dsDNA replicates hauv E Coli thiab tsim ssDNA bearing phages tshiab. Cov phages tshiab no tsis tu ncua tso tawm los ntawm E Coli yam tsis tau tua tus tswv tsev cell. Txawm li cas los xij, tus kab mob ua rau kev loj hlob ntawm E Coli. dsDNA tuaj yeem muab rho tawm los ntawm cov kab mob hlwb thiab siv los ua vectors hauv DNA cloning. Lawv hu ua M13 phage vectors. Lawv tuaj yeem siv tau yooj yim thiab siv zoo ib yam li plasmid vectors.
Lub peev xwm ntawm kev kis tus kab mob ntawm cov M13 phages no ua haujlwm zoo rau kev siv los ua tus vector hauv cov noob cloning. Thaum tsim M13 rau hauv vector, ntau lub ntsiab lus yuav tsum tau muab tso rau hauv nws cov genome. Lawv yog, cov noob rau lac repressor (lac I) protein, tus neeg teb xov tooj-proximal cheeb tsam ntawm lac Z gene, lac txhawb nqa thiab ntau qhov chaw cloning (polylinker). Thaum dsDNA ntawm M13 siv los ua vectors, nws tuaj yeem kho raws li plasmid vector. Txawm li cas los xij, kev siv ssDNA M13 muaj qhov zoo hauv DNA sequencing thiab site-directed mutagenesis.
Txij li thaum M13 phage vector dais ntau qhov chaw cloning hauv thaj av lacZ, cov vectors rov ua dua tuaj yeem txheeb xyuas tau yooj yim los ntawm kev tshuaj ntsuam xyuas xiav / dawb ntawm agar daim hlau uas muaj IPTG thiab X-Gal. Blue plaques uas ua rau ntawm daim hlau tsis muaj cov phages recombined. Yog li ntawd, phages nrog inserts tuaj yeem xaiv rau lub hom phiaj cloning.
Daim duab 02: Bacteriophage M13
YAC thiab M13 Phage Vector yog dab tsi?
YAC vs M13 Phage Vector |
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| YAC yog ib qho kev tsim kho caj ces chromosome uas muaj peev xwm nqa tau ib ntu loj ntawm DNA txawv teb chaws thiab rov ua dua nyob rau hauv cov poov xab hlwb. | M13 phage vector yog tus kab mob vector tsim los ntawm bacteriophage M13 uas yog siv los ntxig txawv DNA rau hauv E Coli. |
| YACs tau tsim los clone cov tawg loj ntawm genomic DNA rau hauv cov poov xab. | M13 phage vectors yog siv los ntxig cov DNA txawv teb chaws rau hauv E Coli. |
| Insert Length | |
| YACs tuaj yeem muaj megabase-loj genomic inserts (1000 kb - 2000 kb). | Qhov loj ntawm cov ntxig yog li 1,500 bps. |
| YAC DNA yog qhov nyuaj rau kev ua kom huv si thiab yuav tsum muaj qhov siab siab rau tsim YAC vector system. | Nws tshwm sim ntawm cyclic photosynthetic electron saw. |
| kev ruaj ntseg | |
| YAC tsis ruaj khov. | M13 phages tau yooj yim muab rho tawm. |
| Enzymes yog cov molecules loj dua. | M13 phages ruaj khov dua YAC. |
Summary – YAC vs M13 Phage Vector
YAC yog ib qho kev tsim kho vector system siv ib cheeb tsam tshwj xeeb ntawm cov poov xab chromosome los ntxig cov khoom siv caj ces rau cov poov xab. M13 phage vector yog vector system muab los ntawm bacteriophage M 13 uas siv E coli ua cov kab mob hauv lub cev. Qhov no yog qhov txawv ntawm YAC thiab M13 Phage Vector. Ob qho tib si muaj txiaj ntsig zoo hauv kev sib txuas DNA thev naus laus zis thiab cov noob qoob loo.
