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PMID: 6218500 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Replication of lambda dv plasmid in vitro promoted by purified lambda O and P proteins.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 79 ·No. 24 ·1982-12-00 ·Pages 7639-43

Tsurimoto T, Matsubara K

Abstract

An in vitro system for replication of lambda dv plasmid DNA has been constructed. This system consists of an ammonium sulfate fraction from Escherichia coli extract, exogenously added purified lambda O and P proteins, and lambda dv DNA in closed circular form. More than 85% of the added template DNA replicated semiconservatively. In the same system, another plasmid, pBR322, also replicated, but less efficiently than lambda dv. Furthermore, its replication was independent of O and P proteins. Inhibitors of DNA gyrase entirely blocked the replication activity, whereas rifampicin, an inhibitor of RNA polymerase, showed a significant effect only when added prior to initiation of the DNA replication. DNA replication was initiated from a region near to or within the four direct repeats in lambda origin (lambda ori) and proceeded bidirectionally, as examined by DNA chain elongation termination with dideoxy CTP. A cloned DNA carrying a 350-base-pair region including the initiation site also initiated replication, dependent on O and P proteins, and its initiation occurred at the same position as with native lambda dv DNA. An A + T-rich structure neighboring the repeats was found to be essential for lambda DNA replication. Regions corresponding to ice and oop were not required for O,P-dependent initiation.

MeSH Terms
Bacteriophage lambda/genetics Base Sequence DNA Replication DNA, Viral/genetics Escherichia coli/genetics Plasmids Viral Proteins/genetics
Chemicals
DNA, Viral Viral Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Tsurimoto T
Matsubara K
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29 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.79.24.7639
Published
1982-12-00
Pages
7639-43
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC347403
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.79.24.7639 PMC Full Text View journal details
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