Quantitative Analysis of the Contributions of Enzyme and DNA to the Structure of λ Integrative Recombinants
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Excerpt
The topology of site-specific recombination has been studied most effectively with substrates in which both sites reside on the same plasmid. Thus, studies of bacteriophage λ integrative recombination by the phage-encoded Int protein and the host proteins complexed in the integration host factor (IHF) have used a series of plasmids containing both the phage attP site and the bacterial attB site. When the 15-bp identical cores in these two sites form a direct repeat and the substrate is supercoiled, the resolution reaction yields products that are catenated recombinant circles with interlinkings of varying complexity (Mizuuchi et al. 1980). Similarly, for recombination between cores in an inverted orientation, the inversion reaction produces knotted products (Nash and Pollock 1983; Pollock and Nash 1983). In this paper we summarize results on the absolute structure of these products from supercoiled substrates, which will be detailed elsewhere (S. Spengler et al., in prep.), and give...
