Thaler D S, Stahl M M, Stahl F W
Institute of Molecular Biology, University of Oregon, Eugene 97403.
J Mol Biol. 1987 May 5;195(1):75-87. doi: 10.1016/0022-2836(87)90328-7.
The Red recombination pathway of phage lambda is shown to target recombination to double-chain ends of DNA. A double-chain cut, delivered in vivo to only one of two parents participating in a lambda lytic cross by a type II restriction endonuclease, increases the proportion of crossing over in the interval containing the cut compared with other intervals. The stimulating effect of a cut is evident whether replication is inhibited or permitted. Cut stimulation can move away from the initial cut-site, presumably by double-chain degradation. Movement of the stimulating effect of a cut is dependent on the Escherichia coli gene recA when the cross is carried out under conditions that inhibit phage replication. When replication is permitted, all aspects of cut-stimulated recombination are independent of recA. Evidence is presented to show that the reaction that is stimulated by cutting is often non-reciprocal at the molecular level.
噬菌体λ的红色重组途径被证明可将重组靶向到DNA的双链末端。通过II型限制性内切酶在体内仅对参与λ裂解杂交的两个亲本之一进行双链切割,与其他区间相比,增加了包含切割位点的区间内的交叉比例。无论复制被抑制还是允许,切割的刺激作用都是明显的。切割刺激可以从初始切割位点移开,推测是通过双链降解。当在抑制噬菌体复制的条件下进行杂交时,切割刺激作用的移动依赖于大肠杆菌基因recA。当允许复制时,切割刺激的重组的所有方面都独立于recA。有证据表明,由切割刺激的反应在分子水平上通常是非互惠的。