Stahl F W, Fox M S, Faulds D, Stahl M M
Institute of Molecular Biology, University of Oregon, Eugene 97403-1229.
Genetics. 1990 Jul;125(3):463-74. doi: 10.1093/genetics/125.3.463.
In phage lambda, when DNA replication is blocked, recombination mediated by the Red pathway occurs only near the double-chain break site, cos, that defines the termini of the virion chromosome. The recombinants initiated by cos contain newly synthesized DNA near cos, in amount corresponding to a few percent of the length of lambda. A restriction enzyme cut delivered to one parent far from cos results in elevated recombination near the restriction site. Recombinants induced by this cut have a similarly small amount of DNA synthesis in these replication-blocked crosses. When restriction cuts are introduced in the presence of normal amounts of all of the DNA replication enzymes, many of the resulting recombinants still enjoy, at most, a small amount of DNA synthesis associated with the exchange event. Thus, these experiments fail to support the previously considered possibility that Red-mediated recombination in lambda proceeds largely through a break-copy pathway.
在λ噬菌体中,当DNA复制受阻时,由Red途径介导的重组仅在双链断裂位点cos附近发生,cos定义了病毒体染色体的末端。由cos引发的重组体在cos附近含有新合成的DNA,其数量相当于λ长度的百分之几。在远离cos的一个亲本中进行的限制性内切酶切割会导致限制性位点附近的重组增加。在这些复制受阻的杂交中,由这种切割诱导的重组体同样只有少量的DNA合成。当在存在正常量的所有DNA复制酶的情况下引入限制性切割时,许多产生的重组体在大多数情况下最多仍只经历与交换事件相关的少量DNA合成。因此,这些实验无法支持之前所认为的λ噬菌体中Red介导的重组主要通过断裂-复制途径进行的可能性。