Cloning of bacterial DNA replication genes in bacteriophage lambda |
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| Authors: | Lee Rowen Joan A. Kobori Stewart Scherer |
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| Affiliation: | (1) Department of Biochemistry, Stanford University School of Medicine, 94305 Stanford, CA, USA;(2) Present address: Division of Biology, California Institute of Technology, 91125 Pasadena, CA, USA;(3) Present address: Division of Chemistry, California Institute of Technology, 91125 Pasadena, CA, USA |
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| Abstract: | Summary Recombinant lambda phages containing the genes for dnaZ protein (the  subunit of DNA polymerse III holoenzyme), primase (dnaG protein) and dnaC protein from Escherichi coli and Salmonella typhimurium were isolated. Each gene cloned from S. typhimurium has extensive DNA sequence homology to the corresponding E. coli gene. Clones selected by complementation of a dnaA temperature-sensitive mutant appear similar to other isolated suppressors of dnaA (Projan and Wechsler 1981). Derivatives of each cloned fragment suitable for overproduction of the protein were constructed. Of those tested, only the phage containing the E. coli dnaZ gene resulted in significant overproduction.Abbreviations DTT dithiothreitol - Ec Escherichia coli - EDTA ethylene diamine tetra acetic acid - kb kilobase 1,000 bases or base-pairs - moi multiplicity of infection - pol I E. coli DNA polymerase I - pol III holoenzyme E. coli DNA polymerase III holoenzyme - pri dnaG, primase-coding gene - SSB single-strand binding protein - St Salmonella typhimurium - sup gene coding for suppressor - ts temperature-sensitive |
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