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PMID: 6262716 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Enzymatic synthesis, ligation, and restriction of DNA containing deoxy-4-thiothymidine.

Nucleic acids research ·Vol. 9 ·No. 4 ·1981-02-25 ·Pages 753-67

Hofer B, Köster H

Abstract

Phage fd RF I DNA1 about 90% substituted by deoxy-4-thiothymidine (s4Td) in the codogenic strand was synthesized by the simultaneous actions of DNA polymerase I and DNA ligase. While the rate of DNA synthesis was considerably reduced, the yield the rate of DNA synthesis was considerably reduced, the yield was not affected in the presence of s4TdTP. The conversion of RF II to RF I DNA by DNA ligase was even improved. This effect seems to be related with an altered ratio of affinity of polymerase and ligase for the s4Td-containing substrate. The presence of the base analogue in the DNA was verified independently by chromatographic and spectroscopic methods. The modified genome could be cleaved by restriction endonucleases Hpa II (C/CGG)d and Taq I (T/CGA)d. A number of the fragments produced showed altered mobilities under the conditions of polyacrylamide gel electrophoresis.

MeSH Terms
Bacteriophages DNA Ligases/metabolism DNA Polymerase I/metabolism DNA Replication DNA Restriction Enzymes/metabolism DNA, Single-Stranded/metabolism DNA, Viral/metabolism Electrophoresis, Agar Gel Escherichia coli Phosphoric Diester Hydrolases/metabolism Spectrophotometry, Ultraviolet Thymidine/analogs & derivatives,metabolism
Chemicals
DNA, Single-Stranded DNA, Viral deoxy-4-thiothymidine DNA Polymerase I DNA Restriction Enzymes Phosphoric Diester Hydrolases DNA Ligases Thymidine
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Hofer B
Köster H
References (34)
34 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/9.4.753
Published
1981-02-25
Pages
753-67
Language
English
Region
England
NLM ID
0411011
PMCID
PMC326710
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/9.4.753 PMC Full Text View journal details
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