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Characterization of an improved in vitro DNA replication system for Escherichia coli plasmids

Conrad, Susan E. and Campbell, Judith L. (1979) Characterization of an improved in vitro DNA replication system for Escherichia coli plasmids. Nucleic Acids Research, 6 (10). pp. 3289-3304. ISSN 0305-1048.

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A modified in vitro replication system has been characterized and used to catalogue the host proteins required for the replication of plasmid RSF1030. These extracts differ from systems described previously in that endogenous DNA is removed. Replication in vitro therefore requires an exogenouos RSF1030 template, even when extracts are prepared from cells carrying endogenous RSF1030. Synthesis in the in vitro system faithfully mimics in vivo replication with respect to the products synthesized, effects of specific inhibitors, and requirements for RNA polymerase and DNA polymerase I. In addition, we find that proteins encoded by dnaB, dnaC, dnaG, dnaI, dnaP and polC (DNA polymerase III), are required for in vitro plasmid synthesis. The product of dnaA is not required. Extracts prepared from E. coli mutants deficient in in vitro replication can be complemented by addition of purified proteins or of extracts carrying the wild type protein.

Item Type:Article
Additional Information:Copyright © 1979 Oxford University Press. Received March 20, 1979. This investigation was supported by grants from the United States Public Health Service: RR07003 and GM 25508. S.E.C. was supported by United States Public Health Service Training Grant No. 5 T32 GM 07616.
Record Number:CaltechAUTHORS:CONnar79
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:4057
Deposited By: Tony Diaz
Deposited On:26 Jul 2006
Last Modified:26 Dec 2012 08:57

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