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Isolation and characterization of lambda pleu bacteriophages.

M G Davis, J M Calvo

    Journal of Bacteriology
    |February 1, 1977
    PubMed
    Summary
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    Researchers engineered bacteriophage lambda to carry the entire leucine operon from Escherichia coli. This allows for the study of leucine biosynthesis regulation and gene expression in bacteria.

    Area of Science:

    • Molecular Biology
    • Bacteriology
    • Genetics

    Background:

    • Escherichia coli leucine operon is essential for leucine biosynthesis.
    • Phage lambda insertion into the leucine operon disrupts enzyme synthesis.
    • Transducing phages can carry bacterial genes.

    Purpose of the Study:

    • To construct and characterize a phage lambda carrying the entire E. coli leucine operon.
    • To investigate the regulation and expression of the leucine operon using engineered phages.
    • To analyze the genetic structure of the leucine operon within phage DNA.

    Main Methods:

    • Heat induction of lysogenic E. coli strains.
    • Construction of recombinant phages via genetic crosses.
    • Analysis of enzyme levels and gene expression.

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  • Restriction endonuclease digestion and heteroduplex analysis of phage DNA.

    • Main Results:

    • Two types of leucine-transducing phages (lambda pleu9 and lambda pleu13) were generated.
    • A phage (lambda K2) carrying the complete leucine operon was constructed.
    • Lysogens with lambda K2 showed functional leucine operon components (leuO, leuP).
    • E. coli leucine-specific mRNA hybridized to the heavy strand of lambda K2.
    • The leucine operon was mapped to a specific DNA fragment (lambdaG4EcoRI-B).

    Conclusions:

    • Engineered phage lambda can carry and express the E. coli leucine operon.
    • This system provides a tool for studying leucine biosynthesis regulation.
    • The genetic organization of the leucine operon within the phage DNA was elucidated.