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枯草芽孢杆菌recM区域的分子克隆、遗传特征分析及DNA序列分析

Molecular cloning, genetic characterization and DNA sequence analysis of the recM region of Bacillus subtilis.

作者信息

Alonso J C, Shirahige K, Ogasawara N

机构信息

Max-Planck-Institut für molekulare Genetik, Berlin, FRG.

出版信息

Nucleic Acids Res. 1990 Dec 11;18(23):6771-7. doi: 10.1093/nar/18.23.6771.

Abstract

In Bacillus subtilis the recM gene, whose product is associated with DNA repair and recombination, has been located between the dnaX and rrnA genes. The recM gene has been cloned and analyzed. Analysis of the nucleotide sequence (3.741-kilobase) around recM revealed five open reading frames (orf). We have assigned recM and dnaX to two of this orf, given the gene order dnaX-orf107-recM-orf74-orf87. The organization of genes of the dnaX-orf107-recM region resembles the organization of genes in the dnaX-orf12-recR region of the Escherichia coli chromosome. Proteins of 24.2 and 17.0 kDa would result from translation of the wild type and in vitro truncated recM genes, and radioactive bands of proteins of molecular weights of 24.5 and 17.0 kDa were detected by the use of the T7promoter-expression system. The RecM protein contains a potential zinc finger domain for nucleic acid binding and a putative nucleotide binding sequence that is present in many proteins that bind and hydrolyze ATP. Strains, in which the recM gene has been insertionally inactivated, were generated and show a phenotype essentially the same as previously described recM mutants.

摘要

在枯草芽孢杆菌中,其产物与DNA修复和重组相关的recM基因位于dnaX和rrnA基因之间。recM基因已被克隆并分析。对recM周围核苷酸序列(3.741千碱基)的分析揭示了五个开放阅读框(orf)。鉴于基因顺序为dnaX-orf107-recM-orf74-orf87,我们已将recM和dnaX指定到其中两个orf。dnaX-orf107-recM区域的基因组织类似于大肠杆菌染色体中dnaX-orf12-recR区域的基因组织。野生型和体外截短的recM基因翻译会产生24.2 kDa和17.0 kDa的蛋白质,通过使用T7启动子表达系统检测到分子量为24.5 kDa和17.0 kDa蛋白质的放射性条带。RecM蛋白包含一个潜在的用于核酸结合的锌指结构域和一个推定的核苷酸结合序列,该序列存在于许多结合并水解ATP的蛋白质中。构建了recM基因已被插入失活的菌株,其表型与先前描述的recM突变体基本相同。

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