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鼠伤寒沙门氏菌LT2的mutL和mutS基因产物的鉴定与特性分析。

Identification and characterization of the mutL and mutS gene products of Salmonella typhimurium LT2.

作者信息

Pang P P, Lundberg A S, Walker G C

出版信息

J Bacteriol. 1985 Sep;163(3):1007-15. doi: 10.1128/jb.163.3.1007-1015.1985.

Abstract

The gene products of the mutL and mutS loci play essential roles in the dam-directed mismatch repair in both Salmonella typhimurium LT2 and Escherichia coli K-12. Mutations in these genes result in a spontaneous mutator phenotype. We have cloned the mutL and mutS genes from S. typhimurium by generating mutL- and mutS-specific probes from an S. typhimurium mutL::Tn10 and an mutS::Tn10 strain and using these to screen an S. typhimurium library. Both the mutL and mutS genes from S. typhimurium were able to complement E. coli mutL and mutS strains, respectively. By a combination of Tn1000 insertion mutagenesis and the maxicell technique, the products of the mutL and mutS genes were shown to have molecular weights of 70,000 and 98,000 respectively. A phi (mutL'-lacZ+) gene fusion was constructed; no change in the expression of the fusion could be detected by treatment with DNA-damaging agents. In crude extracts, the MutS protein binds single-stranded DNA, but not double-stranded DNA, with high affinity.

摘要

mutL和mutS基因座的基因产物在鼠伤寒沙门氏菌LT2和大肠杆菌K-12的dam指导的错配修复中起着至关重要的作用。这些基因的突变会导致自发的突变体表型。我们通过从鼠伤寒沙门氏菌mutL::Tn10和mutS::Tn10菌株中产生mutL和mutS特异性探针,并使用这些探针筛选鼠伤寒沙门氏菌文库,克隆了鼠伤寒沙门氏菌的mutL和mutS基因。鼠伤寒沙门氏菌的mutL和mutS基因分别能够互补大肠杆菌的mutL和mutS菌株。通过Tn1000插入诱变和大细胞技术相结合,显示mutL和mutS基因的产物分子量分别为70,000和98,000。构建了一个phi(mutL'-lacZ+)基因融合体;用DNA损伤剂处理后,未检测到融合体表达的变化。在粗提取物中,MutS蛋白以高亲和力结合单链DNA,但不结合双链DNA。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d28b/219232/8e9e1197d1e9/jbacter00220-0207-a.jpg

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