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大肠杆菌中涉及大异源序列的同源重组。

Homologous recombination involving a large heterology in Escherichia coli.

作者信息

Yamamoto K, Takahashi N, Yoshikura H, Kobayashi I

机构信息

Department of Bacteriology, Faculty of Medicine, University of Tokyo, Japan.

出版信息

Genetics. 1988 Aug;119(4):759-69. doi: 10.1093/genetics/119.4.759.

Abstract

Recombination between two different deletion alleles of a gene (neo) for neomycin and kanamycin resistance was studied in an Escherichia coli sbcA- recB-C- strain. The two homologous regions were in an inverted orientation on the same plasmid molecule. Kanamycin-resistant plasmids were selected and analyzed. The rate of recombination to form kanamycin-resistant plasmids was decreased by mutations in the recE, recF and recJ genes, but was not decreased by a mutation in the recA gene. It was found that these plasmids often possessed one wild-type kanamycin-resistant allele (neo+) while the other neo allele was still in its original (deletion) form. Among kanamycin-resistant plasmids with one wild-type and one parental allele it was often found that the region between the inverted repeats had been flipped (turned around) with respect to sites outside the inverted repeats. These results were interpreted as follows. Gene conversion, analogous to gene conversion in eukaryotic meiosis, is responsible for a unidirectional transfer of information from one neo deletion allele to the other. The flipping of the region between the inverted repeats is interpreted as analogous to the crossing over associated with gene conversion in eukaryotic meiosis. In contrast with a rec+ strain, these products cannot be explained by two rounds of reciprocal crossing over involving a dimeric form as an intermediate. In the accompanying paper we present evidence that gene conversion by double-strand gap repair takes place in the same E. coli strain.

摘要

在一株大肠杆菌sbcA- recB-C-菌株中,研究了对新霉素和卡那霉素具有抗性的基因(neo)的两个不同缺失等位基因之间的重组。两个同源区域在同一质粒分子上呈反向排列。选择并分析了卡那霉素抗性质粒。形成卡那霉素抗性质粒的重组率因recE、recF和recJ基因的突变而降低,但不因recA基因的突变而降低。发现这些质粒通常拥有一个野生型卡那霉素抗性等位基因(neo+),而另一个neo等位基因仍处于其原始(缺失)形式。在具有一个野生型和一个亲本等位基因的卡那霉素抗性质粒中,经常发现反向重复序列之间的区域相对于反向重复序列之外的位点发生了翻转(旋转)。这些结果的解释如下。类似于真核生物减数分裂中的基因转换,基因转换负责将信息从一个neo缺失等位基因单向转移到另一个。反向重复序列之间区域的翻转被解释为类似于真核生物减数分裂中与基因转换相关的交叉。与rec+菌株不同,这些产物不能用涉及二聚体形式作为中间体的两轮相互交叉来解释。在随附的论文中,我们提供了证据表明双链缺口修复的基因转换发生在同一大肠杆菌菌株中。

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