[The cloning and expression of the RNAse structural gene of Bacillus intermedius].

E B Chernokal'skaia, E R Romakhina, N P Balaban, M R Sharipova, V I Vershinina, I B Leshchinskaia
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Abstract

The structure gene of extracellular alkaline ribonuclease Bacillus intermedius (binase) has been cloned in E. coli cells in composition of pMT 316 plasmid carrying the inhibitor gene (barstar of barnase--binase structure homologue. The possibility to use such vector has been proved during the barstar action on binase catalytic activity. Using biochemical immunochemical analysis the expression of binase gene in E. coli cells has been confirmed. The recombinant clone E. coli which contains both plasmids simultaneously--carrying gene for barster and for benase has been produced. The given vector is suggested to be used for cloning of inhibitor gene to obtain a viable producer of alkaline intracellular ribonuclease.

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[中间芽孢杆菌RNAse结构基因的克隆与表达]。
胞外碱性核糖核酸酶中间芽孢杆菌(二酶)的结构基因在大肠杆菌细胞中克隆,其结构基因由携带二酶抑制基因(二酶-二酶结构同源物)的pMT 316质粒组成。在barstar对二酶催化活性的作用中,已经证明了使用这种载体的可能性。利用生化免疫化学分析证实了二酶基因在大肠杆菌细胞中的表达。制备了同时携带barster基因和benase基因的重组大肠杆菌克隆。建议利用该载体克隆抑制基因,获得碱性细胞内核糖核酸酶的活性产物。
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