Tests of the Double-Strand-Break Repair Model for Red-Mediated Recombination of Phage and Plasmid dv

David S. Thaler ¹, Mary M. Stahl ¹, and Franklin W. Stahl ¹

¹ Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403

The double-strand-break repair (DSBR) model was formulated to account for various aspects of yeast mitotic and meiotic recombination. In this study three features of the DSBR model are tested for Red-mediated recombination between phage and dv, a plasmid that is perfectly homologous to about 10% of . The results support the applicability of the DSBR model to 's Red system: (1) Creating a double-strand-break (DSB) within the region of homology shared by phage and plasmid increases their genetic interaction by about 20-fold. A DSB outside the region of shared homology has no such effect. (2) Both patches, i.e., simple marker rescue, and splices, i.e., co-integration of the phage and plasmid, are stimulated by a DSB in the region of shared homology. (3) Co-integrants harbor a duplication of the region of shared homology. Among co-integrants that were formed by the creation of a DSB, there is a preferential loss of whichever allele was in cis to a utilized cut site. The DSBR model as originally formulated involves the isomerization and cleavage of Holliday junctions to resolve the canonical intermediate. We propose as an alternative mechanism that a topoisomerase can resolve the canonical DSBR intermediate.

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