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PMID: 6956881 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Functional origin of replication of pT181 plasmid DNA is contained within a 168-base-pair segment.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 79 ·No. 15 ·1982-08-00 ·Pages 4580-4

Khan SA, Adler GK, Novick RP

Abstract

We have used a recently developed in vitro replication system from Staphylococcus aureus to determine the origin and direction of replication of pT181 plasmid DNA. The origin was located to within 168 base pairs by two methods: (i) sequential labeling of restriction endonuclease fragments after synchronous initiation in vitro in the presence of various amounts of dideoxy-TTP and (ii) by constructing in vitro deletions of pT181 DNA close to the origin of replication and testing for their ability to replicate in vitro pT181 plasmid was found to replicate unidirectionally and anticlockwise, as the map is conventionally drawn. The nucleotide sequence of the region containing the origin of replication has been determined and found to be partially or entirely contained within the coding sequence for the repC protein, which is uniquely required for pT181 plasmid replication. Preliminary evidence suggesting that pT181 replicates by a rolling circle mechanism is discussed.

MeSH Terms
Bacterial Proteins/metabolism Base Sequence Chromosome Mapping DNA Replication DNA, Bacterial Nucleic Acid Conformation Plasmids Replicon Staphylococcus aureus
Chemicals
Bacterial Proteins DNA, Bacterial RepC protein, Staphylococcus aureus
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Khan S A
Adler G K
Novick R P
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32 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.79.15.4580
Published
1982-08-00
Pages
4580-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC346718
Subset
IM
Databases
GENBANK
J01764, J01765
External Links
PubMed source DOI: 10.1073/pnas.79.15.4580 PMC Full Text View journal details
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