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Cloning of bacterial DNA replication genes in bacteriophage λ

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Summary

Recombinant lambda phages containing the genes for dnaZ protein (the γ 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.

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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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Author information

Author notes

  1. Joan A. Kobori

    Present address: Division of Biology, California Institute of Technology, 91125, Pasadena, CA, USA

  2. Stewart Scherer

    Present address: Division of Chemistry, California Institute of Technology, 91125, Pasadena, CA, USA

Authors and Affiliations

  1. Department of Biochemistry, Stanford University School of Medicine, 94305, Stanford, CA, USA

    Lee Rowen, Joan A. Kobori & Stewart Scherer

Authors
  1. Lee Rowen
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  2. Joan A. Kobori
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  3. Stewart Scherer
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Additional information

Communicated by H.W. Boyer

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Rowen, L., Kobori, J.A. & Scherer, S. Cloning of bacterial DNA replication genes in bacteriophage λ. Molec Gen Genet 187, 501–509 (1982). https://doi.org/10.1007/BF00332635

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  • DOI: https://doi.org/10.1007/BF00332635

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