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Genetics, Vol 147, 961-977, Copyright © 1997
INVESTIGATIONS |
Annealing vs. Invasion in Phage {lambda} Recombination
M. M. Stahl, L. Thomason, A. R. Poteete, T. Tarkowski, A. Kuzminov and F. W. Stahl
Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229.
Genetic recombination catalyzed by {lambda}'s Red pathway was studied in rec(+) and recA mutant bacteria by examining both intracellular {lambda} DNA and mature progeny particles. Recombination of nonreplicating phage chromosomes was induced by double-strand breaks delivered at unique sites in vivo. In rec(+) cells, cutting only one chromosome gave nearly maximal stimulation of recombination; the recombinants formed contained relatively short hybrid regions, suggesting strand invasion. In contrast, in recA mutant cells, cutting the two parental chromosomes at non-allelic sites was required for maximal stimulation; the recombinants formed tended to be hybrid over the entire region between the two cuts, implying strand annealing. We conclude that, in the absence of RecA and the presence of non-allelic DNA ends, the Red pathway of {lambda} catalyzes recombination primarily by annealing.
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