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大肠杆菌质粒中Tn5倒置事件的分析。

Analysis of Tn5 inversion events in Escherichia coli plasmids.

作者信息

Weber P C

机构信息

Pennsylvania State University College of Medicine, Milton S. Hershey Medical Center, Department of Microbiology and Immunology, Hershey 17033, USA.

出版信息

Mol Gen Genet. 1995 Aug 30;248(4):459-70. doi: 10.1007/BF02191646.

Abstract

The ability of the bacterial transposon Tn5 to undergo sequence inversion in Rec+ Escherichia coli cells as a result of recombination between its duplicated IS50 elements was examined using specially designed plasmid constructs. Surprisingly, recombination events in the IS50 elements that led to crossover and therefore Tn5 inversion could be detected at a frequency of only 10(-5). This was approximately an order of magnitude lower than the frequency of IS50 recombination that led to conversion events (i.e. non-reciprocal recombination) without crossover, and at least two orders of magnitude lower than the frequency of intermolecular recombination between IS50 elements on two different plasmids. These rare conversion and inversion events in Tn5 appeared to be due to intramolecular recombination and not simply to multiple rounds of reciprocal crossing over, since the heterodimeric intermediates that would be generated during the latter process could be readily isolated but were shown to yield a completely different set of plasmid products upon resolution.

摘要

利用特殊设计的质粒构建体,研究了细菌转座子Tn5在Rec⁺大肠杆菌细胞中因其重复的IS50元件之间发生重组而进行序列倒位的能力。令人惊讶的是,导致交叉从而导致Tn5倒位的IS50元件中的重组事件,仅能以10⁻⁵的频率被检测到。这比导致无交叉的转换事件(即非相互重组)的IS50重组频率低约一个数量级,并且比两个不同质粒上的IS50元件之间的分子间重组频率至少低两个数量级。Tn5中这些罕见的转换和倒位事件似乎是由于分子内重组,而不仅仅是多轮相互交叉,因为在后一过程中会产生的异二聚体中间体可以很容易地分离出来,但在拆分后显示会产生完全不同的一组质粒产物。

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