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通过构建操纵子融合体对大肠杆菌uvrC基因调控元件的分析。

Analysis of the regulatory elements of the Escherichia coli uvrC gene by construction of operon fusions.

作者信息

Forster J W, Strike P

机构信息

Department of Agricultural Botany, University College of Wales, Dyfed, UK.

出版信息

Mol Gen Genet. 1988 Mar;211(3):531-7. doi: 10.1007/BF00425712.

Abstract

The regulatory region of the Escherichia coli uvrC gene has been analysed by the subcloning of appropriate restriction fragments into the promoter probe vector pPV502. A series of plasmids carrying operon fusions to the gene for chloramphenicol acetyltransferase (cat) has been constructed. Three promoters capable of controlling uvrC have been identified (P1, P2 and P3), the majority of transcription being derived from the most distal of these promoters (P1). Transcription termination apparently plays some role in the control of the gene through premature termination of the P1-, but not the P2- or P3-derived transcripts. In addition, a promoter acting in the opposite direction to uvrC transcription has been detected. The activity of each of the promoters has been assayed under normal and SOS-inducing conditions. The uvrC gene is not apparently under the control of the recA-lexA regulatory circuit, unlike uvrA and uvrB. A series of recombinant plasmids carrying a 1.9 kb Bg/II fragment encoding most of the uvrC gene has been constructed. The properties of these plasmids suggest that the six amino acids at the carboxy-terminus of the uvrC gene product are not critical for DNA repair activity.

摘要

通过将合适的限制性片段亚克隆到启动子探针载体pPV502中,对大肠杆菌uvrC基因的调控区域进行了分析。构建了一系列携带与氯霉素乙酰转移酶(cat)基因的操纵子融合体的质粒。已鉴定出三个能够控制uvrC的启动子(P1、P2和P3),大部分转录来自这些启动子中最远端的启动子(P1)。转录终止显然通过P1衍生转录本的提前终止在该基因的调控中发挥了一定作用,但对P2或P3衍生的转录本没有影响。此外,还检测到一个与uvrC转录方向相反的启动子。在正常和SOS诱导条件下对每个启动子的活性进行了测定。与uvrA和uvrB不同,uvrC基因显然不受recA-lexA调控回路的控制。构建了一系列携带编码uvrC基因大部分的1.9 kb Bg/II片段的重组质粒。这些质粒的特性表明,uvrC基因产物羧基末端的六个氨基酸对DNA修复活性并不关键。

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