Becker E C, Zhou H, Meyer R J
Department of Microbiology, University of Texas at Austin, USA.
J Bacteriol. 1996 Aug;178(16):4870-6. doi: 10.1128/jb.178.16.4870-4876.1996.
The origin of replication of the plasmid R1162 contains an initiation site for the synthesis of each DNA strand. When one of these sites (oriL) is deleted, synthesis on the corresponding strand is no longer initiated efficiently in vitro by the R1162-encoded replication proteins, and the plasmid is no longer stably maintained in the cell. However, in vivo the two strands of the plasmid duplex molecule are active at a similar level as templates for DNA synthesis, and newly synthesized copies of each strand are incorporated into daughter molecules at a similar rate. No secondary, strong initiation sites on the delta oriL strand were detected in the region of the origin. The delta oriL plasmid induces the SOS response, and this is important for plasmid maintenance even in a recombination-proficient strain. Our results indicate that an SOS-induced host system can maintain an R1162 derivative lacking one of its initiation sites.
质粒R1162的复制起点包含每条DNA链合成的起始位点。当其中一个位点(oriL)缺失时,在体外由R1162编码的复制蛋白不再能有效地起始相应链上的合成,并且该质粒在细胞中不再稳定维持。然而,在体内,质粒双链分子的两条链作为DNA合成模板的活性水平相似,并且每条链新合成的拷贝以相似的速率掺入子代分子中。在起点区域未检测到δoriL链上有二级强起始位点。δoriL质粒诱导SOS应答,这对于质粒维持很重要,即使在重组功能正常的菌株中也是如此。我们的结果表明,SOS诱导的宿主系统可以维持缺少其一个起始位点的R1162衍生物。