Chernokal'skaia E B, Romakhina E R, Balaban N P, Sharipova M R, Vershinina V I, Leshchinskaia I B
Nauchnye Doki Vyss Shkoly Biol Nauki. 1992(2):45-50.
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.
中间芽孢杆菌胞外碱性核糖核酸酶(binase)的结构基因已在大肠杆菌细胞中克隆,该细胞含有携带抑制剂基因(barnase的barstar——binase结构同源物)的pMT 316质粒。在barstar对binase催化活性的作用过程中,已证明使用这种载体的可能性。通过生化免疫化学分析,已证实binase基因在大肠杆菌细胞中的表达。已产生同时含有两种质粒的重组大肠杆菌克隆——携带barster基因和benase基因。建议使用给定的载体克隆抑制剂基因,以获得碱性细胞内核糖核酸酶的可行生产者。