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鞭毛甲基杆菌recA基因在大肠杆菌中的克隆、测序及表达

Cloning, sequence and expression in Escherichia coli of the Methylobacillus flagellatum recA gene.

作者信息

Gomelsky M, Gak E, Chistoserdov A, Bolotin A, Tsygankov Y D

机构信息

Institute of Genetics and Selection of Industrial Microorganisms, Moscow, U.S.S.R.

出版信息

Gene. 1990 Sep 28;94(1):69-75. doi: 10.1016/0378-1119(90)90469-8.

Abstract

By means of interspecific complementation of an Escherichia coli recA- mutation with phasmids containing a gene bank from an obligate methylotroph, Methylobacillus flagellatum (Mf), the recA+ gene from this bacterium was identified. When expressed in an E. coli recA- host, it can function in recombination, DNA repair, and prophage induction. The nucleotide sequence of the gene has been determined. The coding region consists of 1032 bp specifying 344 amino acids. The deduced RecA protein structure shows a striking homology with RecA from other bacteria, except for the C-terminal region and some residues which were proposed to be responsible for the coprotease ability of RecA proteins. The region preceding the recA-Mf gene start codon has no SOS box--the LexA repressor binding site. Expression of the recA-Mf gene in E. coli proved to be DNA-damage independent.

摘要

通过用含有专性甲基营养菌鞭毛甲基杆菌(Mf)基因文库的噬菌粒对大肠杆菌recA - 突变进行种间互补,鉴定出了该细菌的recA + 基因。当在大肠杆菌recA - 宿主中表达时,它可在重组、DNA修复和原噬菌体诱导中发挥作用。已确定了该基因的核苷酸序列。编码区由1032个碱基对组成,指定344个氨基酸。推导的RecA蛋白结构与其他细菌的RecA显示出显著的同源性,但C末端区域和一些被认为负责RecA蛋白共蛋白酶能力的残基除外。recA - Mf基因起始密码子之前的区域没有SOS框——LexA阻遏物结合位点。recA - Mf基因在大肠杆菌中的表达被证明与DNA损伤无关。

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