Ligation of double-stranded and single-stranded [Oligo(dT)] DNA by vaccinia virus DNA ligase
Vaccinia virus DNA ligase has been expressed in <em>Escherichia coli</em>, purified, and biochemically characterized. The enzyme ligates double-stranded (ds) DNA substrates with either cohesive or blunt-end termini and the latter reaction is stimulated by PEG. Vaccinia virus DNA ligase c...
| Autori principali: | , , |
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| Natura: | Journal article |
| Lingua: | English |
| Pubblicazione: |
Elsevier
1996
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| Soggetti: |
| _version_ | 1826261288972451840 |
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| author | Odell, M Kerr, S Smith, G |
| author_facet | Odell, M Kerr, S Smith, G |
| author_sort | Odell, M |
| collection | OXFORD |
| description | Vaccinia virus DNA ligase has been expressed in <em>Escherichia coli</em>, purified, and biochemically characterized. The enzyme ligates double-stranded (ds) DNA substrates with either cohesive or blunt-end termini and the latter reaction is stimulated by PEG. Vaccinia virus DNA ligase can also ligate oligo(dT) when annealed to either a poly(dA) or a poly(rA) backbone and, remarkably, free oligo(dT). This ligation of a single-stranded (ss) substrate is unique among eukaryotic DNA ligases. The enzyme requires high ATP concentrations with a <i>K<sub>m</sub></i> for the overall ligation of a ssDNA substrate of 0.8 m<i>M</i>. The salt, divalent cation, temperature, and pH requirements of the enzyme for the optimal ligation of ss and ds substrate are described. |
| first_indexed | 2024-03-06T19:19:06Z |
| format | Journal article |
| id | oxford-uuid:196eaad0-e524-421a-855b-eb3781d880df |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T19:19:06Z |
| publishDate | 1996 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | oxford-uuid:196eaad0-e524-421a-855b-eb3781d880df2022-03-26T10:49:04ZLigation of double-stranded and single-stranded [Oligo(dT)] DNA by vaccinia virus DNA ligaseJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:196eaad0-e524-421a-855b-eb3781d880dfVirusesGenetics (medical sciences)EnglishOxford University Research Archive - ValetElsevier1996Odell, MKerr, SSmith, GVaccinia virus DNA ligase has been expressed in <em>Escherichia coli</em>, purified, and biochemically characterized. The enzyme ligates double-stranded (ds) DNA substrates with either cohesive or blunt-end termini and the latter reaction is stimulated by PEG. Vaccinia virus DNA ligase can also ligate oligo(dT) when annealed to either a poly(dA) or a poly(rA) backbone and, remarkably, free oligo(dT). This ligation of a single-stranded (ss) substrate is unique among eukaryotic DNA ligases. The enzyme requires high ATP concentrations with a <i>K<sub>m</sub></i> for the overall ligation of a ssDNA substrate of 0.8 m<i>M</i>. The salt, divalent cation, temperature, and pH requirements of the enzyme for the optimal ligation of ss and ds substrate are described. |
| spellingShingle | Viruses Genetics (medical sciences) Odell, M Kerr, S Smith, G Ligation of double-stranded and single-stranded [Oligo(dT)] DNA by vaccinia virus DNA ligase |
| title | Ligation of double-stranded and single-stranded [Oligo(dT)] DNA by vaccinia virus DNA ligase |
| title_full | Ligation of double-stranded and single-stranded [Oligo(dT)] DNA by vaccinia virus DNA ligase |
| title_fullStr | Ligation of double-stranded and single-stranded [Oligo(dT)] DNA by vaccinia virus DNA ligase |
| title_full_unstemmed | Ligation of double-stranded and single-stranded [Oligo(dT)] DNA by vaccinia virus DNA ligase |
| title_short | Ligation of double-stranded and single-stranded [Oligo(dT)] DNA by vaccinia virus DNA ligase |
| title_sort | ligation of double stranded and single stranded oligo dt dna by vaccinia virus dna ligase |
| topic | Viruses Genetics (medical sciences) |
| work_keys_str_mv | AT odellm ligationofdoublestrandedandsinglestrandedoligodtdnabyvacciniavirusdnaligase AT kerrs ligationofdoublestrandedandsinglestrandedoligodtdnabyvacciniavirusdnaligase AT smithg ligationofdoublestrandedandsinglestrandedoligodtdnabyvacciniavirusdnaligase |