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Ligation of double-stranded and single-stranded [Oligo(dT)] DNA by vaccinia virus DNA ligase

Abstract:
Vaccinia virus DNA ligase has been expressed in Escherichia coli, 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 Km for the overall ligation of a ssDNA substrate of 0.8 mM. The salt, divalent cation, temperature, and pH requirements of the enzyme for the optimal ligation of ss and ds substrate are described.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1006/viro.1996.0358

Authors

More by this author
Institution:
University of Oxford
Division:
MSD
Department:
Pathology Dunn School
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
Pathology Dunn School
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
Pathology Dunn School
Role:
Author


Publisher:
Elsevier
Journal:
Virology More from this journal
Volume:
221
Issue:
1
Pages:
120–129
Publication date:
1996-07-01
Edition:
Publisher's version
DOI:
ISSN:
0042-6822


Language:
English
Subjects:
UUID:
uuid:196eaad0-e524-421a-855b-eb3781d880df
Local pid:
ora:10722
Deposit date:
2015-03-25
ARK identifier:

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