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Related Experiment Videos

Complementation between temperature-sensitive and deletion mutants of reovirus.

D A Spandidos, A F Graham

    Journal of Virology
    |December 1, 1975
    PubMed
    Summary

    Reovirus temperature-sensitive (ts) mutants were tested for complementation using a plaque assay. Class C mutants were unique, neither complementing nor being complemented, leading to their assignment to the L1 viral genome segment.

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    Area of Science:

    • Virology
    • Molecular Biology
    • Genetics

    Background:

    • Temperature-sensitive (ts) mutants are crucial tools for studying viral gene function.
    • Reovirus, a segmented dsRNA virus, exhibits complex genetic interactions.
    • Previous studies indicated low levels of complementation between reovirus ts mutants.

    Purpose of the Study:

    • To definitively characterize complementation among reovirus ts mutants.
    • To assign reovirus ts mutations to specific viral genome segments.
    • To investigate the genetic role of the L1 segment.

    Main Methods:

    • Development of a sensitive plaque assay for reovirus complementation.
    • Mixed infection of cell monolayers with defective reovirions and ts mutants.
    • Complementation analysis at a non-permissive temperature (39°C).

    Main Results:

    • Classes A, B, D, F, and G reovirus ts mutants showed complementation with defective viruses.
    • Class C and E mutants did not complement with defective viruses.
    • Class C mutants neither complemented nor were complemented by defective virions, while Class G complemented defective virions.

    Conclusions:

    • The class C reovirus ts mutation was assigned to the L1 viral genome segment.
    • This assignment is based on the unique lack of complementation observed for class C mutants.
    • The study provides a refined method for reovirus genetic analysis.

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