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大肠杆菌中影响ColE1维持的herC突变体的分离与鉴定

Isolation and characterization of herC, a mutation of Escherichia coli affecting maintenance of ColE1.

作者信息

Kawakami K, Naito S, Inoue N, Nakamura Y, Ikeda H, Uchida H

机构信息

Institute of Medical Science, University of Tokyo, Japan.

出版信息

Mol Gen Genet. 1989 Nov;219(3):333-40. doi: 10.1007/BF00259604.

Abstract

Two modes of ColE1 DNA replication are known, one dependent on RNase H, and the other RNase H independent. The cer114 mutant of the ColE1 replicon is defective in both modes and carries a single base pair alteration 95 bp upstream of the replication origin. An Escherichia coli mutant which restored maintenance of the cer114 replicon was isolated. This host suppressor mutant is defective in RNase H and carries a herC mutation located at 62 min of the E. coli chromosome. The herC mutation is recessive to its wild-type allele and supports maintenance of the mutant replicon in the absence of RNase H. The herC mutation alone conferred cold-sensitive growth, suggesting that the herC gene product is essential for cell growth. The 1832 bp E. coli DNA fragment, containing the wild-type allele of the herC mutation, was cloned and an open reading frame for the HerC protein was determined.

摘要

已知ColE1 DNA复制有两种模式,一种依赖核糖核酸酶H,另一种不依赖核糖核酸酶H。ColE1复制子的cer114突变体在这两种模式下均有缺陷,且在复制起点上游95 bp处有一个单碱基对改变。分离出了一个能恢复cer114复制子稳定性的大肠杆菌突变体。这个宿主抑制突变体在核糖核酸酶H方面有缺陷,且在大肠杆菌染色体62分钟处有一个herC突变。herC突变对其野生型等位基因是隐性的,并且在没有核糖核酸酶H的情况下支持突变复制子的稳定性。单独的herC突变赋予了冷敏感生长特性,这表明herC基因产物对细胞生长至关重要。克隆了包含herC突变野生型等位基因的1832 bp大肠杆菌DNA片段,并确定了HerC蛋白的开放阅读框。

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