Stahl M M, Thomason L, Poteete A R, Tarkowski T, Kuzminov A, Stahl F W
Institute of Molecular Biology, University of Oregon, Eugene 97403-1229, USA.
Genetics. 1997 Nov;147(3):961-77. doi: 10.1093/genetics/147.3.961.
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.
通过检测细胞内λ DNA和成熟子代颗粒,在rec⁺和recA突变细菌中研究了由λ噬菌体的Red途径催化的基因重组。通过在体内独特位点产生的双链断裂诱导非复制型噬菌体染色体的重组。在rec⁺细胞中,仅切割一条染色体就能几乎最大程度地刺激重组;形成的重组体包含相对较短的杂交区域,表明有链侵入。相比之下,在recA突变细胞中,需要在非等位位点切割两条亲代染色体才能实现最大程度的刺激;形成的重组体倾向于在两次切割之间的整个区域杂交,这意味着链退火。我们得出结论,在没有RecA且存在非等位DNA末端的情况下,λ噬菌体的Red途径主要通过退火催化重组。