Qhov Tseem Ceeb - T4 vs T7 DNA Ligase
DNA ligase yog ib qho tseem ceeb enzyme siv hauv molecular biology cov txheej txheem. Nws ua haujlwm raws li cov kua nplaum molecular los koom nrog nucleotides los ntawm kev tsim phosphodiester bonds ntawm nucleotides. Cov phosphodiester bonds yog tsim los ntawm 3 'hydroxyl kawg ntawm qab zib moiety thiab 5' kawg phosphate pawg. DNA ligase yog qhov tseem ceeb hauv cov txheej txheem rov ua dua los koom nrog Okazaki fragments ntawm lub lagging strand thiab thaum lub sij hawm kho DNA mechanisms thiab hauv vitro cloning thwmsim los koom nrog cov noob xav tau kev txaus siab rau vector genome. Muaj ob lub ntsiab DNA ligases siv tam sim no los ntawm molecular biologists: T4 thiab T7 DNA ligases. T4 DNA ligase yog ib qho ntawm thawj enzymes raug cais tawm ntawm T4 bacteriophage. T7 DNA ligase, uas yog cov protein me me, yog ib qho enzyme cais los ntawm T7 bacteriophage. Qhov no yog qhov sib txawv tseem ceeb ntawm T4 thiab T7 DNA ligases.
DNA Ligase yog dab tsi?
DNA ligation tshwm sim hauv peb-kauj ruam cov tshuaj tiv thaiv. Thawj kauj ruam yog kev tawm tsam nucleophilic ntawm alpha-phosphorus ntawm ATP tsim kom muaj covalent enzyme-adenylate nruab nrab uas AMP txuas nrog lub davhlau ya nyob twg-chain nitrogen ntawm lysine. Hauv theem ob, kev tawm tsam nucleophilic tshwm sim ntawm phosphorus ntawm enzyme-adenylate los ntawm 5'-phosphate-terminated DNA substrate, tso lysine thiab tsim DNA-adenylate. Hauv cov kauj ruam kawg, DNA-adenylate raug tawm tsam los ntawm 3'- OH ntawm lwm DNA strand, tso tawm AMP thiab koom nrog cov polynucleotides.
T4 DNA Ligase yog dab tsi?
T4 Ligase yog thawj ligase uas raug cais thiab txheeb xyuas los ntawm Meselson, Weigle, thiab Kellenberger thiab yog thawj zaug lag luam ligase tsim tawm. T4 DNA ligase yog ATP nyob ntawm enzyme uas catalyzes phosphodiester daim ntawv cog lus. Nws yog ib leeg polypeptide saw nrog 487 amino acid residues thiab muaj qhov hnyav molecular txog 77 kDa. Qhov zoo tshaj pH rau nws cov dej num nyob hauv thaj tsam ntawm 7.5 - 8.
Daim duab 01: T4 DNA Ligase
T4 DNA Ligase, raws li nws lub npe qhia, muaj nyob hauv bacteriophage T4 thiab raug cais tawm thaum phage tsim cov enzyme thaum kis kab mob E. coli. Lub enzyme ligates ob txoj hlua DNA duplexes nrog qhov kawg kawg thiab cov tawg tawg yuav tsum nyob ze rau ib leeg.
T7 DNA Ligase yog dab tsi?
T7 ligase yog 41 kDa protein uas tsim los ntawm T7 Bacteriophage thiab cais los ntawm kev kis kab mob E. coli hlwb. Lub luag haujlwm tseem ceeb ntawm T7 ligase yog txhawm rau txhim kho DNA duplexes los ntawm kev sib txuas ntawm phosphodiester thiab koom nrog cov seem ntawm DNA nyob ze thaum kev sim cloning. Qhov zoo tshaj pH rau T7 DNA ligase nyob hauv thaj tsam ntawm 7.0 - 7.2. T7 ligase yog nyob ntawm ATP. T7 DNA ligases kuj muaj peev xwm ntawm kev sib xyaw DNA thiab RNA hybrids, ua rau muaj kev txawv txav txawv txav thaum lub sijhawm hloov pauv; Txawm li cas los xij, qhov tshwm sim no tseem nyob hauv kev tshawb fawb los ntawm kev tshawb fawb hauv vitro.
Dab tsi zoo sib xws ntawm T4 thiab T7 DNA Ligase?
- T4 thiab T7 DNA Ligase koom nrog kev ligation cov tshuaj tiv thaiv los ntawm annealing DNA duplexes los ntawm phosphodiester bond tsim ntawm 3 'OH kawg thiab 5' phosphate kawg ntawm DNA fragment.
- Ob lub enzymes ligate blunt kawg DNA duplexes uas nyob ze rau ib leeg.
- Ob lub enzymes yog ATP nyob ntawm.
- Cov enzymes no raug cais los ntawm kev kis kab mob E. coli nrog cov kab mob tshwj xeeb.
- Kev siv ntawm ob lub enzymes suav nrog:
- Molecular cloning - kom ligate cov noob ntawm kev txaus siab rau vector
- kho DNA - kho ob txoj hlua khi
- YIn vitro DNA replication
- Ligase chain reaction
Qhov txawv ntawm T4 thiab T7 DNA Ligase yog dab tsi?
T4 vs T7 DNA Ligase |
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| T4 DNA Ligase hom DNA ligase cais tawm ntawm cov kab mob bacteriophage T4 thiab koom nrog ligating cov DNA uas nyob ib sab ntawm phosphodiester daim ntawv cog lus. | T7 DNA Ligase hom DNA ligase cais tawm los ntawm cov kab mob bacteriophage T7 thiab koom nrog ligating cov DNA uas nyob ib sab ntawm phosphodiester daim ntawv cog lus. |
| Size ntawm Protein | |
| Loj ntawm T4 DNA ligase yog 77 kDa - loj dua piv rau T7 DNA ligase. | Loj T7 DNA ligase yog 41 kDa. |
| zoo pH | |
| pH ntau yam uas ua haujlwm T4 DNA ligase yog 7.5 - 8.0. | pH ntau yam uas ua haujlwm T7 DNA ligase yog 7.0 - 7.2. |
Summary – T4 vs T7 DNA Ligase
DNA ligases yog cov chav kawm tseem ceeb ntawm cov enzymes koom nrog hauv molecular cloning los tsim cov DNA recombinant molecules uas muaj cov noob paj. Ligation cov tshuaj tiv thaiv yog lub zog siv cov tshuaj tiv thaiv thiab muaj cov kev tshawb fawb tshiab tab tom txhim kho qhov tseeb ntawm kev hloov kho ligases muaj. T4 thiab T7 DNA ligases yog ob hom tseem ceeb DNA ligase enzymes siv nyob rau hauv molecular biological txuj ci. T7 DNA ligase kuj yog qhov tshwm sim ntawm kev tshawb fawb uas tau ua raws li thawj ligase cais los ntawm T4 tus kab mob E. coli. T7 yog portrayed ntau efficiency thiab raug nyob rau hauv nqa tawm nws muaj nuj nqi vim nws me me. Qhov no yog qhov txawv ntawm T4 thiab T7 ligase.
Download PDF Version of T4 vs T7 DNA Ligase
Koj tuaj yeem rub tawm PDF version ntawm kab lus no thiab siv nws rau offline hom phiaj raws li cov ntawv sau tseg. Thov download PDF version ntawm no Qhov txawv nruab nrab ntawm T4 thiab T7 Ligase.
