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大肠杆菌ruv基因的遗传分析与分子克隆

Genetic analysis and molecular cloning of the Escherichia coli ruv gene.

作者信息

Shurvinton C E, Lloyd R G, Benson F E, Attfield P V

出版信息

Mol Gen Genet. 1984;194(1-2):322-9. doi: 10.1007/BF00383535.

Abstract

The genetic organisation of the ruv gene, a component of the SOS system for DNA repair and recombination in Escherichia coli, was investigated. New point mutations as well as insertions and deletions were generated using transposon Tn10 inserted in eda as a linked marker for site specific mutagenesis, or directly as a mutagen. The mutations were ordered with respect to one another and previously isolated ruv alleles by means of transductional crosses. The direction of chromosome mobilization from ruv ::Mud( ApRlac ) strains carrying F42lac + established that ruv is transcribed in a counterclockwise direction. Recombinant lambda phages able to restore UV resistance to ruv mutants were identified,and the ruv + region was subcloned into a low copy number plasmid. The ruv + plasmid was able to correct the extreme radiation sensitivity and recombination deficiency of ruv recBC sbcB strains.

摘要

对大肠杆菌中DNA修复和重组的SOS系统的一个组成部分ruv基因的遗传组织进行了研究。利用插入eda中的转座子Tn10作为位点特异性诱变的连锁标记,或直接作为诱变剂,产生了新的点突变以及插入和缺失。通过转导杂交将这些突变相互排序,并与先前分离的ruv等位基因进行排序。从携带F42lac+的ruv::Mud(ApRlac)菌株进行染色体移动的方向确定ruv是以逆时针方向转录的。鉴定出能够恢复ruv突变体紫外线抗性的重组λ噬菌体,并将ruv+区域亚克隆到低拷贝数质粒中。ruv+质粒能够纠正ruv recBC sbcB菌株的极端辐射敏感性和重组缺陷。

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