AI Article Synopsis
- Murine polyomavirus mutants are created for research and therapeutic uses, but current methods for preparing them are often complex and yield inconsistent results.
- The authors developed a straightforward Cre/loxP recombination method that generates intact polyomavirus genomes from recombinant plasmids directly in living cells.
- Their study found that while one of the recombinant constructs successfully produced infectious virus, the other did not, demonstrating the effectiveness of their method.
Article Abstract
Murine polyomavirus mutants are frequently produced for experimental as well as therapy purposes. Commonly used methods for preparation of mutant viral genomes from recombinant vectors are laborious and give variable yields and quality. We describe an efficient and reproducible Cre/loxP-mediated recombination system that generates polyomavirus genomes from recombinant plasmid in vivo. We designed and constructed two variants of recombinant vectors containing the wild-type polyomavirus genome flanked by loxP homologous sites. The loxP sites were introduced either into the intronic region of early genes or between the two poly(A) signal sites of convergent transcriptional units. After cotransfection of the recombinant plasmids with the Cre-expressing vector into mouse 3T6 cells, we obtained infectious virus from the genome variant containing loxP site in the intronic region, but we failed to isolate any infectious virus from the viral genome containing loxP site between poly(A) signals. We show that the Cre/loxP-based method of polyomavirus production is simple, expedient, and reproducible and works with satisfactory efficiency.
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| http://dx.doi.org/10.1016/j.virusres.2013.05.016 | DOI Listing |
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