Document Detail

Replication of mini-P1 plasmid DNA in vitro requires two initiation proteins, encoded by the repA gene of phage P1 and the dnaA gene of Escherichia coli.
MedLine Citation:
PMID:  3035546     Owner:  NLM     Status:  MEDLINE    
We have developed an in vitro DNA-replication system that replicates exogenously added mini-P1 plasmid DNA. The system consists of purified P1 RepA protein and a partially purified mixture of Escherichia coli replication proteins. It is essentially the same as that described for the replication of oriC plasmid DNA [Fuller, R.S., Kaguni, J.M. & Kornberg, A. (1981) Proc. Natl. Acad. Sci. USA 78, 7370-7374]. Mini-P1 DNA replication requires the E. coli DnaA initiation protein in addition to the P1 RepA initiation protein. The reaction is inhibited by rifampicin, novobiocin, and antibody to DnaB, suggesting the involvement of RNA polymerase, DNA gyrase, and DnaB protein. Replication is initiated in the region of the P1 origin of replication and proceeds unidirectionally as determined by electron microscopy. Thus, the in vitro system mimics the essential features of mini-P1 replication as suggested by genetic studies.
S H Wickner; D K Chattoraj
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  84     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  1987 Jun 
Date Detail:
Created Date:  1987-07-08     Completed Date:  1987-07-08     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  3668-72     Citation Subset:  IM    
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MeSH Terms
Bacterial Proteins / genetics*
Coliphages / genetics*
DNA Helicases*
DNA Replication*
DNA, Bacterial / ultrastructure
DNA-Binding Proteins*
Escherichia coli / genetics*
Genes, Bacterial*
Genes, Viral*
Viral Proteins / genetics*
Reg. No./Substance:
0/Bacterial Proteins; 0/DNA, Bacterial; 0/DNA-Binding Proteins; 0/Proteins; 0/Trans-Activators; 0/Viral Proteins; 0/replication initiator protein; EC 3.6.1.-/DNA Helicases

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