Characterization of an improved in vitro DNA replication system for Escherichia coli plasmids.
AUTOR(ES)
Conrad, S E
RESUMO
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. 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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=327934Documentos Relacionados
- Overproduction of Escherichia coli replication proteins by the use of runaway-replication plasmids.
- Novel densitometric method for endonuclease analysis of Escherichia coli DNA samples containing multiple plasmids.
- DNA transcription and repressor binding affect deletion formation in Escherichia coli plasmids.
- Deletion hot spots in chimeric Escherichia coli plasmids.
- Termination sites T1 and T2 from the Escherichia coli chromosome inhibit DNA replication in ColE1-derived plasmids.