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dnaB analog function associated with plasmid R100drd-1.

A A Potter, V N Iyer

    Journal of Bacteriology
    |March 1, 1983
    PubMed
    Summary
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    Plasmid R100 derivatives suppress dnaB gene temperature sensitivity in E. coli. An R100dnaB analog function was identified, distinct from phage P1 and P7, supporting bacteriophage lambda growth.

    Area of Science:

    • Microbiology
    • Molecular Biology
    • Genetics

    Background:

    • The dnaB gene is essential for DNA replication in Escherichia coli K-12.
    • Temperature-sensitive dnaB mutations can lead to cell death at restrictive temperatures.
    • Plasmids are extrachromosomal DNA elements that can carry genes conferring new traits.

    Purpose of the Study:

    • To investigate the ability of plasmid R100 derivatives to suppress temperature-sensitive dnaB mutations in E. coli.
    • To identify and characterize a potential dnaB analog function encoded by plasmid R100.
    • To compare the functional properties of the R100-encoded dnaB analog with those of bacteriophages P1 and P7.

    Main Methods:

    • Complementation assays using E. coli strains with different dnaB alleles.

    Related Experiment Videos

  • Testing the ability of plasmid R100 derivatives to rescue dnaB mutations.
  • Evaluating the support of bacteriophage lambda growth in a dnaB mutant background.
  • Thermosensitivity assays for the dnaB analog function.

    • Main Results:

    • Plasmid R100 and its derivatives suppressed temperature sensitivity of various dnaB alleles.
    • R100drd-l and pAR132 rescued a dnaB266(Am) mutation without amber suppressors, indicating an R100-encoded dnaB analog function.
    • The R100-encoded dnaB analog supported bacteriophage lambda growth in a dnaB266(Am) strain, differentiating it from P1 and P7 dnaB analogs.
    • The dnaB analog function encoded by R100drd-l was found to be thermosensitive.

    Conclusions:

    • Plasmid R100 encodes a functional dnaB analog that can complement dnaB defects in E. coli.
    • This R100-derived dnaB analog exhibits distinct properties compared to those from bacteriophages P1 and P7.
    • The structural gene for the R100 dnaB analog is located within the EcoRI fragment D of R100drd-l.