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Related Experiment Videos

Methods for identification of recombinants of phage lambda.

B Sanzey, O Mercereau, T Ternynck

    Proceedings of the National Academy of Sciences of the United States of America
    |October 1, 1976
    PubMed
    Summary

    Two methods enable screening of phage plaques for specific DNA sequences or antigens. These techniques utilize lambda and lambdalac phages for efficient identification of phage-encoded genetic material and proteins.

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    Area of Science:

    • Molecular Biology
    • Virology
    • Immunology

    Background:

    • Phage display technology is crucial for identifying specific DNA sequences and antigens.
    • Efficient screening methods are needed for large-scale phage plaque analysis.
    • Lambda and lambdalac phages are widely used model systems in molecular biology.

    Purpose of the Study:

    • To describe two novel methods for screening phage plaques.
    • To detect specific DNA sequences and antigens within phage plaques.
    • To assess the utility of these methods for identifying lambda transducers.

    Main Methods:

    • Phage plaques transferred to nitrocellulose filters via desiccation.
    • Detection of lac DNA sequence using hybridization with radioactive lac mRNA.

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  • Detection of Beta-Galactosidase antigen using immune serum and enzyme-coupled antibodies.

    • Main Results:

    • Successful detection of specific DNA sequences (lac) in phage plaques.
    • Successful detection of specific antigens (Beta-Galactosidase) in phage plaques.
    • Demonstrated applicability to lambda and lambdalac phage systems.

    Conclusions:

    • The described methods provide efficient screening for phage-borne DNA sequences and antigens.
    • These techniques are valuable for identifying lambda transducers, including those from in vitro recombination.
    • The methods enhance the capabilities of phage display and genetic screening.