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枯草芽孢杆菌核糖核酸酶III基因:克隆、该基因在大肠杆菌中的功能以及构建核糖核酸酶III基因座改变的枯草芽孢杆菌菌株

Bacillus subtilis RNase III gene: cloning, function of the gene in Escherichia coli, and construction of Bacillus subtilis strains with altered rnc loci.

作者信息

Wang W, Bechhofer D H

机构信息

Department of Biochemistry, Mount Sinai School of Medicine of the City University of New York, New York 10029, USA.

出版信息

J Bacteriol. 1997 Dec;179(23):7379-85. doi: 10.1128/jb.179.23.7379-7385.1997.

Abstract

The rnc gene of Bacillus subtilis, which has 36% amino acid identity with the gene that encodes Escherichia coli RNase III endonuclease, was cloned in E. coli and shown by functional assays to encode B. subtilis RNase III (Bs-RNase III). The cloned B. subtilis rnc gene could complement an E. coli rnc strain that is deficient in rRNA processing, suggesting that Bs-RNase III is involved in rRNA processing in B. subtilis. Attempts to construct a B. subtilis rnc null mutant were unsuccessful, but a strain was constructed in which only a carboxy-terminal truncated version of Bs-RNase III was expressed. The truncated Bs-RNase III showed virtually no activity in vitro but was active in vivo. Analysis of expression of a copy of the rnc gene integrated at the amy locus and transcribed from a p(spac) promoter suggested that expression of the B. subtilis rnc is under regulatory control.

摘要

枯草芽孢杆菌的rnc基因与编码大肠杆菌核糖核酸酶III内切核酸酶的基因具有36%的氨基酸同一性,该基因在大肠杆菌中克隆,并通过功能测定表明其编码枯草芽孢杆菌核糖核酸酶III(Bs - 核糖核酸酶III)。克隆的枯草芽孢杆菌rnc基因可以补充rRNA加工缺陷的大肠杆菌rnc菌株,这表明Bs - 核糖核酸酶III参与枯草芽孢杆菌的rRNA加工。构建枯草芽孢杆菌rnc缺失突变体的尝试未成功,但构建了一个仅表达Bs - 核糖核酸酶III羧基末端截短版本的菌株。截短的Bs - 核糖核酸酶III在体外几乎没有活性,但在体内有活性。对整合在amy位点并从p(spac)启动子转录的rnc基因拷贝的表达分析表明,枯草芽孢杆菌rnc的表达受调控控制。

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