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Related Experiment Videos

Intracistronic complementation in the simian virus 40 A gene.

J Tornow, C N Cole

    Proceedings of the National Academy of Sciences of the United States of America
    |October 1, 1983
    PubMed
    Summary
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    Molecular biology of the cell·1998

    Researchers created simian virus 40 (SV40) mutants with deletions in the large tumor antigen gene. In-phase deletions complemented a nonviable mutant, suggesting distinct functional domains within the SV40 T antigen.

    Area of Science:

    • Molecular Biology
    • Virology
    • Genetics

    Background:

    • Simian virus 40 (SV40) is a well-characterized DNA tumor virus.
    • The SV40 large tumor (T) antigen is crucial for viral replication and oncogenesis.
    • Understanding T antigen domains is key to deciphering its functions.

    Purpose of the Study:

    • To investigate the functional domains of the SV40 T antigen.
    • To correlate specific mutations within the T antigen gene with functional complementation.
    • To analyze the impact of in-phase versus frameshift deletions on T antigen activity.

    Main Methods:

    • Construction of eight SV40 mutants with small deletions (3-20 base pairs) in the A gene at specific map units.
    • Isolation of mutants with both in-phase and frameshift deletions.

    Related Experiment Videos

  • Transfection of monkey kidney cells to assess plaque formation and DNA replication.
  • Complementation assays involving cotransfection with a nonviable mutant (dlA2459).

    • Main Results:

    • Mutants with frameshift deletions failed to replicate or form plaques.
    • A mutant with an in-phase deletion (dlA2459) was replication-positive.
    • Four mutants with in-phase deletions complemented dlA2459, enabling plaque formation.
    • Mutants with frameshift deletions did not complement dlA2459.

    Conclusions:

    • The results suggest that in-phase deletions within the SV40 T antigen gene can restore function through complementation.
    • Frameshift deletions lead to a loss of function, indicating their critical impact.
    • The defect in dlA2459 likely represents a distinct functional domain of the SV40 T antigen, separable from other domains affected by frameshift mutations.