Kong D, Masker W
Department of Biochemistry and Molecular Biology, Temple University School of Medicine, Philadelphia, Pennsylvania 19140.
J Bacteriol. 1994 Oct;176(19):5904-11. doi: 10.1128/jb.176.19.5904-5911.1994.
An in vitro system based on extracts of Escherichia coli infected with bacteriophage T7 was used to study genetic deletions between directly repeated sequences. The frequency of deletion was highest under conditions in which the DNA was actively replicating. Deletion frequency increased markedly with the length of the direct repeat both in vitro and in vivo. When a T7 gene was interrupted by 93 bp of nonsense sequence flanked by 20-bp direct repeats, the region between the repeats was deleted in about 1 out of every 1,600 genomes during each round of replication. Very similar values were found for deletion frequency in vivo and in vitro. The deletion frequency was essentially unaffected by a recA mutation in the host. When a double-strand break was placed between the repeats, repair of this strand break was often accompanied by the deletion of the DNA between the direct repeats, suggesting that break rejoining could contribute to deletion during in vitro DNA replication.
基于感染噬菌体T7的大肠杆菌提取物构建的体外系统,用于研究直接重复序列之间的基因缺失。在DNA进行活跃复制的条件下,缺失频率最高。在体外和体内,缺失频率均随直接重复序列长度的增加而显著升高。当一个T7基因被一段93 bp的无义序列中断,该无义序列两侧为20 bp的直接重复序列时,在每一轮复制过程中,每1600个基因组中约有1个基因组的重复序列之间的区域会被删除。在体内和体外发现的缺失频率非常相似。宿主中的recA突变对缺失频率基本没有影响。当在重复序列之间产生双链断裂时,该链断裂的修复常常伴随着直接重复序列之间DNA的缺失,这表明断裂重接可能在体外DNA复制过程中导致缺失。