Kong D, Masker W
Department of Biochemistry and Molecular Biology, Temple University School of Medicine, Philadelphia, Pennsylvania 19140.
J Biol Chem. 1993 Apr 15;268(11):7721-7.
An in vitro system based upon extracts of bacteriophage T7 infected Escherichia coli was used to study genetic deletions and to examine the importance of DNA replication in the deletion process. When T7 genomes with gene 1.3 inactivated by a 43-bp insert of random sequence DNA bracketed by 11-bp direct repeats were replicated in vitro the inserts were deleted with a frequency of about 10(-5) deletions per genome replication. Under conditions where deletion could take place only by recombination between direct repeats on distinct DNA molecules deletion frequency was at least an order of magnitude lower. These data demonstrate the utility of the in vitro system as a means for examining deletion mechanisms and underscore the importance of DNA replication in deletions.
基于噬菌体T7感染的大肠杆菌提取物构建的体外系统,用于研究基因缺失,并检验DNA复制在缺失过程中的重要性。当带有被11个碱基对的直接重复序列包围的随机序列DNA的43个碱基对插入片段而使基因1.3失活的T7基因组在体外复制时,每个基因组复制中插入片段的缺失频率约为10^(-5)。在缺失只能通过不同DNA分子上的直接重复序列之间的重组发生的条件下,缺失频率至少低一个数量级。这些数据证明了该体外系统作为研究缺失机制手段的实用性,并强调了DNA复制在缺失中的重要性。