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Related Experiment Videos

Multiple origins and circular structures in replicating T5 bacteriophage DNA.

G J Bourguignon, T K Sweeney, H Delius

    Journal of Virology
    |April 1, 1976
    PubMed
    Summary

    Replication of T5 phage DNA initiates bidirectionally from multiple internal origins. Both linear and circular DNA molecules exhibit complex replication patterns, including theta and sigma forms, indicating a dynamic process.

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    Area of Science:

    • Molecular Biology
    • Virology
    • Genetics

    Background:

    • T5 phage DNA replication is a complex process.
    • Understanding replication origins is crucial for viral DNA synthesis.

    Purpose of the Study:

    • To investigate the origins and patterns of T5 phage DNA replication.
    • To characterize the structures of replicating T5 DNA molecules.

    Main Methods:

    • Gentle isolation of T5 phage DNA using NaI density gradient centrifugation.
    • Examination of DNA molecules via electron microscopy.

    Main Results:

    • Early replication shows linear T5 DNA with multiple "eye-loops" originating bidirectionally from internal sites.
    • A primary origin is near the T5 genome center, not at nicks.

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  • Later stages reveal circular T5 DNA (replicating and non-replicating) and various replication intermediates (theta, sigma forms).

    • Conclusions:

    • T5 phage DNA replication initiates from multiple internal origins.
    • Replication proceeds bidirectionally, with complex intermediate structures observed.
    • Circularization of DNA occurs during later stages of replication.