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Related Experiment Videos

In vitro assembly of bacteriophage Lambda heads.

D Kaiser, T Masuda

    Proceedings of the National Academy of Sciences of the United States of America
    |January 1, 1973
    PubMed
    Summary

    Lambda phage assembly was studied in cell-free extracts. Researchers found that head gene products from genes A and E act early in the assembly pathway, preceding genes B, C, and D.

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    Area of Science:

    • Bacteriophage biology
    • Molecular genetics
    • Virology

    Background:

    • Bacteriophages, like lambda phage, are viruses that infect bacteria.
    • Understanding phage assembly is crucial for molecular biology and genetic engineering.
    • Lambda phage assembly involves complex interactions of structural proteins and DNA packaging.

    Purpose of the Study:

    • To investigate the assembly pathway of plaque-forming lambda phage particles in cell-free extracts.
    • To identify the temporal order of head gene products during phage assembly.
    • To elucidate the role of specific head genes (A, B, C, D, E) in the assembly process.

    Main Methods:

    • Cell-free extract complementation assays using lambda lysogens with amber mutations in head genes.
    • In vitro assembly experiments involving the addition of purified phage DNA to cell-free extracts.
    • Characterization of assembled plaque-formers for DNase resistance, antibody neutralization, adsorption specificity, and buoyant density.

    Main Results:

    • Plaque-forming particles were successfully assembled in cell-free extracts, possessing properties of normal phage.
    • Complementation analysis revealed that mutants in genes B, C, or D could donate DNA, but mutants in A or E could not.
    • Complementation occurred only when at least one extract acted as a DNA donor, indicating specific roles in assembly.

    Conclusions:

    • The results suggest a tentative pathway for lambda phage head assembly.
    • Products of genes A and E appear to act early in the assembly process.
    • Products of genes B, C, and D function later in the head assembly pathway.

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