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Plaque assay for lambda transducing phage carrying the E. coli metB gene.

J R Johnson, C A Liljestrand

    Molecular & General Genetics : MGG
    |January 1, 1983
    PubMed
    Summary

    A new halo plaque assay detects lambda transducing phages with functional metB genes in Escherichia coli K12. This method uses phage plaques and methionine-limited bacterial lawns to identify gene function via visible growth haloes.

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    Area of Science:

    • Microbiology
    • Molecular Biology
    • Genetics

    Background:

    • Bacterial genetics research relies on identifying functional genes.
    • Lambda phage is a tool for gene transfer (transduction) in bacteria.
    • Detecting specific gene alleles, like metB, is crucial for understanding bacterial metabolism.

    Purpose of the Study:

    • To develop a novel assay for detecting lambda transducing phages.
    • To specifically identify phages carrying functional alleles of the metB gene in Escherichia coli K12.
    • To establish a versatile method applicable to various biosynthetic markers.

    Main Methods:

    • A halo plaque assay was designed using metB- Escherichia coli K12 bacterial lawns.
    • Lawns were supplemented with limiting amounts of methionine.
    • Detection involved observing haloes of bacterial growth around phage plaques, indicating successful transduction and prototrophy.

    Main Results:

    • The halo plaque assay successfully detected nondefective lambda transducing phages carrying functional metB alleles.
    • Characteristic halo morphologies allowed for precise genotype determination.
    • The assay demonstrated effectiveness in identifying prototrophic transductants.

    Conclusions:

    • The developed halo plaque assay is an effective tool for detecting specific gene alleles in bacteria.
    • This technique offers a reliable method for identifying functional metB genes in Escherichia coli K12.
    • The assay's general applicability to biosynthetic markers broadens its utility in microbial genetics.

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