Hastings P J, McGill C, Shafer B, Strathern J N
Department of Genetics, University of Alberta, Edmonton, Canada.
Genetics. 1993 Dec;135(4):973-80. doi: 10.1093/genetics/135.4.973.
Integration of linearized plasmids into yeast chromosomes has been used as a model system for the study of recombination initiated by double-strand breaks. The linearized plasmid DNA recombines efficiently into sequences homologous to the ends of the DNA. This efficient recombination occurs both for the configuration in which the break is in a contiguous region of homology (herein called the ends-in configuration) and for "omega" insertions in which plasmid sequences interrupt a linear region of homology (herein called the ends-out configuration). The requirements for integration of these two configurations are expected to be different. We compared these two processes in a yeast strain containing an ends-in target and an ends-out target for the same cut plasmid. Recovery of ends-in events exceeds ends-out events by two- to threefold. Possible causes for the origin of this small bias are discussed. The lack of an extreme difference in frequency implies that cooperativity between the two ends does not contribute to the efficiency with which cut circular plasmids are integrated. This may also be true for the repair of chromosomal double-strand breaks.
将线性化质粒整合到酵母染色体中已被用作研究由双链断裂引发的重组的模型系统。线性化的质粒DNA能有效地重组到与DNA末端同源的序列中。这种高效重组在断裂位于同源连续区域的构型(在此称为末端向内构型)以及质粒序列中断同源线性区域的“ω”插入(在此称为末端向外构型)中均会发生。预计这两种构型整合的要求会有所不同。我们在一个酵母菌株中比较了这两个过程,该菌株针对同一切割质粒含有一个末端向内靶点和一个末端向外靶点。末端向内事件的回收率比末端向外事件高出两到三倍。讨论了这种微小偏差产生的可能原因。频率上没有极端差异意味着两端之间的协同作用对切割环状质粒的整合效率没有贡献。这对于染色体双链断裂的修复可能也是如此。