Transcription-replication collision increases recombination efficiency between plasmids
Transcription-replication collision increases recombination efficiency between plasmids
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DOI：
10.1016/j.plasmid.2013.10.001
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发表时间：
2013-11-01
期刊：
影响因子：
2.6
通讯作者：
Liu DePei
中科院分区：
文献类型：
--
作者：
Li Jialiang;Chen Feng;Liu DePei
有人提出，在一些生物体中，复制叉的停滞可以诱导同源重组，并且停滞的复制叉可能提供核酸酶靶点，从而为参与双链修复的蛋白质提供底物。在本文中，我们构建了一个具有转录复制碰撞（TRC）潜力的质粒，其中DNA复制和RNA转录同时发生在同一DNA模板上。从理论上讲，转录会阻碍DNA复制，增加同源重组。为了验证这一假设，我们构建了另一个含有同源序列的质粒，除了一些突变位点。将这两种质粒共转染到293T细胞中导致重组频率增加。这两个质粒的比例也影响重组频率。此外，我们发现RAD51的高表达水平，这表明重组率的增加可能是通过同源重组途径。这些结果表明，trc诱导的重组可以修复质粒中的突变基因。(C) 2013爱思唯尔公司版权所有。
It has been proposed that the stalling of the replication forks can induce homologous recombination in several organisms, and that arrested replication forks may offer nuclease targets, thereby providing a substrate for proteins involved in double-strand repair. In this article, we constructed a plasmid with the potential for transcription-replication collision (TRC), in which DNA replication and RNA transcription occur on the same DNA template simultaneously. Theoretically, transcription will impede DNA replication and increase homologous recombination. To validate this hypothesis, another plasmid was constructed that contained a homologous sequence with the exception of some mutated sites. Co-transfection of these two plasmids into 293T cells resulted in increased recombination frequency. The ratio of these two plasmids also affected the recombination frequency. Moreover, we found high expression levels of RAD51, which indicated that the increase in the recombination rate was probably via the homologous recombination pathway. These results indicate that mutant genes in plasmids can be repaired by TRC-induced recombination. (C) 2013 Elsevier Inc. All rights reserved.