Simian virus 40 recombinants are produced at high frequency during infection with genetically mixed oligomeric DNA.
- 1 June 1979
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 76 (6) , 2876-2880
- https://doi.org/10.1073/pnas.76.6.2876
Abstract
Classical approached to analysis of mitotic recombination by use of SV-40 are limited in usefulness because of low frequencies of recombination. To bypass the apparent rate-limiting step in normal SV-40 recombination, oligomeric SV-40 was constructed in vitro by ligation of mixtures of pairs of linear DNA carrying genetically distinct temperature-sensitive mutations. Cultured monkey cells infected with the unfractionated ligation products yielded frequencies of nonparental recombinant progeny that were increased up to 500-fold relative to cells infected with a mixture of the untreated circular molecules. Pairwise corsses were performed with tsB4, tsB8 and tsBC11 DNA, using unfractionated oligomers constructed from linear molecules cleaved by EcoRI or BamHI. In each cross the fraction of progeny with nonparental genotypes was roughly proportional to the physical distances between the mutant sites. These results suggest a random, rather than site-specific, conversion of oligomers to monomers. Somewhat surprisingly, nonligated mixtures of linear tsB4 and tsB8 DNA, created by EcoRI digestion, produced a 40- to 100-fold increase in the frequency of nonparental progeny. Intermolecular associations must occur with fairly high efficiency between these linear molecules.This publication has 26 references indexed in Scilit:
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