PubMed · 1367802
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M J Geisow. 1991. Characterizing recombinant proteins.. https://doi.org/10.1038/nbt1091-921
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We report the rescue of a defective vaccinia virus, forming the basis for a stringent selection protocol to generate replicating recombinant virus without the need for marker cassettes and selection agents. Plaques of recombinant virus could be isolated solely by their ability to grow in wild-type cells normally supporting the growth of vaccinia virus. All growth-competent clones analyzed contained the gene of interest in the intended genomic locus and displayed foreign gene expression to the same levels as was seen with classical recombinants obtained by insertion into the vaccinia virus thymidine kinase locus. The system is based on a defective vaccinia virus, expressing exclusively early genes, termed eVAC-1, and an insertion plasmid vector providing the essential function, the uracil DNA glycosylase gene. In addition, the defective virus is free of selection and color marker genes, thus also representing a basic vector for the generation of defective recombinants.
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The tangle model developed by Ernst and Sumners provides a rigorous framework to study processive DNA recombination. We suggest here a slight modification of that model. The tangle equations become: N(S) = b(1, 1), N(S + M) = b(2, 1), N(S + M + M) = b(5, 2), N(S + M + M + M) = b(8, 5), N(S + M + M + M + M) = b(11, 7), where M is the mechanism tangle, and S is the substrate tangle, that is the sum of O (outside tangle) and P (parent tangle). The advantage of this revisited model is that it faithfully models the fact that the recombination mechanism is the same during each event of recombination. This leads to new solutions for O and P, some of which are interesting from a biological viewpoint.