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Related Experiment Videos

Isolation of coliphage lambda ghosts able to adsorb onto bacterial cells

G Konopa, K Taylor

    Biochimica Et Biophysica Acta
    |August 13, 1975
    PubMed
    Summary

    We explored three methods for producing lambda ghosts. Treatment with EDTA resulted in ghosts that effectively adsorb to bacteria, unlike those from osmotic shock or LiCl methods.

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    Area of Science:

    • Molecular Biology
    • Virology
    • Biochemistry

    Background:

    • Bacteriophage lambda is a key model organism in molecular biology.
    • Bacteriophage lambda ghosts are valuable tools for genetic engineering and research.
    • Efficient production of functional lambda ghosts is crucial for various applications.

    Purpose of the Study:

    • To evaluate and compare three distinct methods for producing lambda ghosts.
    • To identify a method yielding lambda ghosts with optimal adsorption properties.
    • To assess the purity of lambda ghosts produced by different methods.

    Main Methods:

    • Utilized [3H]leucine-labeled bacteriophage lambda.
    • Employed CsCl density gradient sedimentation for purification.
    • Investigated three ghost production techniques: osmotic shock, 5 M LiCl incubation, and ethylenediaminetetraacetic acid (EDTA) treatment.

    Main Results:

    • Ghosts produced via osmotic shock or 5 M LiCl treatment exhibited poor bacterial adsorption.
    • EDTA-treated ghosts, purified by CsCl density gradient sedimentation, retained significant adsorption capabilities.
    • EDTA-treated ghosts were found to be substantially free of both DNA and infectious phage particles.

    Conclusions:

    • Ethylenediaminetetraacetic acid (EDTA) treatment is a superior method for producing functional lambda ghosts.
    • EDTA-treated lambda ghosts maintain essential adsorption properties for bacterial interaction.
    • This method yields highly purified, non-infectious lambda ghost preparations suitable for research.

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