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Related Experiment Videos

Analysis of the phiX DNA replication cycle by electron microscopy.

K Koths, D Dressler

    Proceedings of the National Academy of Sciences of the United States of America
    |February 1, 1978
    PubMed
    Summary

    This study reveals two DNA replication intermediates during phiX virus infection: a rolling circle and a novel partially duplex circle. These findings clarify viral DNA synthesis and polymerization rates.

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    Area of Science:

    • Virology
    • Molecular Biology
    • Genetics

    Background:

    • Understanding viral DNA replication is crucial for antiviral development.
    • The phiX virus life cycle provides a model for studying viral DNA synthesis.

    Purpose of the Study:

    • To elucidate the intracellular DNA forms during the phiX virus life cycle.
    • To characterize the intermediates of viral DNA replication.

    Main Methods:

    • Infection of individual cells with single virions.
    • One-step DNA isolation and purification.
    • Quantitative electron microscopy for DNA form analysis.

    Main Results:

    • Early infection shows rolling circle and novel partially duplex single-stranded circle intermediates.
    • Late infection reveals only rolling circle intermediates (approx. 35 per cell).
    • Net polymerization rate in rolling circles is ~200 nucleotides/sec.

    Conclusions:

    • Viral DNA replication occurs in two asymmetric steps: positive strand synthesis followed by negative strand synthesis.
    • The rolling circle with a single-stranded tail is a key intermediate in phiX DNA replication.
    • Replication intermediates and polymerization rates provide insights into viral genome duplication.

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