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大肠杆菌K12的recN基因座:基因产物的分子分析与鉴定

The recN locus of Escherichia coli K12: molecular analysis and identification of the gene product.

作者信息

Picksley S M, Morton S J, Lloyd R G

出版信息

Mol Gen Genet. 1985;201(2):301-7. doi: 10.1007/BF00425675.

Abstract

The recN gene which is necessary for inducible DNA repair and recombination in Escherichia coli has been cloned into the low copy plasmid vector pHSG415. Analysis of the recombinant plasmid, pSP100, revealed a 5.6 Kb HindIII insert of chromosomal DNA. Transposon inactivation of recN function and analysis of a recN::Mu(Ap lac) fusion located the coding region to a 1.4 Kb region within a 2.1 Kb BglII-AvaI DNA fragment transcribed in a clockwise direction with respect to the chromosome map. The gene product was identified in maxicells as a 60,000 dalton protein. Synthesis of this protein was increased in cells lacking LexA activity or in strains carrying recN cloned into the multicopy vector pBR322. Multiple copies of recN increase resistance to ionizing radiation in recN mutants but reduce the survival of a wild-type strain.

摘要

对大肠杆菌中诱导型DNA修复和重组所必需的recN基因已被克隆到低拷贝质粒载体pHSG415中。对重组质粒pSP100的分析显示,其含有一个5.6 Kb的染色体DNA HindIII插入片段。recN功能的转座子失活以及对recN::Mu(Ap lac)融合体的分析将编码区域定位到一个2.1 Kb BglII - AvaI DNA片段内的1.4 Kb区域,该片段相对于染色体图谱按顺时针方向转录。在大细胞中鉴定出该基因产物是一种60,000道尔顿的蛋白质。在缺乏LexA活性的细胞中或携带克隆到多拷贝载体pBR322中的recN的菌株中,这种蛋白质的合成增加。recN的多个拷贝增加了recN突变体对电离辐射的抗性,但降低了野生型菌株的存活率。

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