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PMID: 3523437 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The involvement of host replication proteins and of specific origin sequences in the in vitro replication of miniplasmid R1 DNA.

Nucleic acids research ·Vol. 14 ·No. 12 ·1986-06-25 ·Pages 4865-79

Ortega S, Lanka E, Diaz R

Abstract

The in vitro replication of R1 miniplasmid promoted by purified preparations of the plasmid encoded RepA protein in cell extracts of E. coli is resistant to rifampicin and can be completely inhibited by antibodies against DnaG, the primase of the cell, as well as by antibodies against proteins DnaB and SSB. R1 replication is abolished in extracts deficient in the DnaA protein. This deficiency is efficiently complemented by purified preparations of the DnaA protein. The in vitro replication of plasmid R1 is also abolished in DnaC deficient extracts and by a 10 bp deletion (nucleotides 1463-1472) within the minimal origin region. These data indicate the requirement of the DnaA, DnaB, DnaC, DnaG and SSB replication proteins of the host, as well as of specific oriR1 sequences for the RepA dependent replication of plasmid R1. The implications of these results for the initiation of R1 replication are discussed.

MeSH Terms
Bacterial Proteins/genetics Cell-Free System DNA Replication DNA, Bacterial/genetics DNA-Binding Proteins/genetics Escherichia coli/genetics In Vitro Techniques Plasmids
Chemicals
Bacterial Proteins DNA, Bacterial DNA-Binding Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ortega S
Lanka E
Diaz R
References (45)
45 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/14.12.4865
Published
1986-06-25
Pages
4865-79
Language
English
Region
England
NLM ID
0411011
PMCID
PMC311497
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/14.12.4865 PMC Full Text View journal details
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