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Published on: January 12, 2015
Site-specific excision of integrated polyoma DNA
Murine Cyp cells excise integrated polyoma virus DNA upon temperature shift-down. This precise excision involves a recombination mechanism, creating chimeric molecules with specific DNA sequence similarities at the crossover site.
Area of Science:
- Molecular Biology
- Virology
- Genetics
Background:
- Cyp cells are murine cells with a thermosensitive polyoma virus genome.
- The viral genome integrates at 39°C and excises/replicates at 33°C.
Purpose of the Study:
- To investigate the mechanism of precise DNA excision in rare Cyp cell subclones.
- To identify the cellular and viral sequences involved in generating chimeric molecules (Rm I).
Main Methods:
- Analysis of DNA sequences at the crossover site in chimeric molecules.
- Comparison of viral and cellular DNA sequences flanking the integration junction.
Main Results:
- Temperature shift-down induces excision and circularization of specific DNA segments in rare Cyp subclones.
- The crossover site exhibits minimal direct homology (1-2 base pairs).
- Extensive homology patches (19-20 base pairs) and oppositely oriented similar sequences (12-14 base pairs) flank the crossover site.
Conclusions:
- A specific recombination mechanism mediates accurate viral DNA excision.
- The observed DNA sequence features suggest a novel mechanism for site-specific recombination.
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