Avakov A S, Bolotin A P, Kolibaba L G, Sorokin A V, Shemiakina T M, Paberit M, Raĭk Kh, Aaviksaar A
Mol Biol (Mosk). 1990 Jul-Aug;24(4):1001-9.
Plasmids pCB20 and pCB22 were used for cloning and expression of the Bac brevis 7882 neutral protease gene in Bac. subtilis cells. The protease-containing fragments of 13 and 14 kb were cloned in pCB20 plasmid based on replication region of Streptococci plasmid pSM19035. Expression of the gene was shown to take place in Bac. subtilis. Application of vegetative promoters of the previously identified expression unit EU19035 greatly increases the expression of the protease in Bac. subtilis. Bac. subtilis cells, expressing the gene of Bac. brevis neutral protease, do not sporulate, are considerably larger than the cells which do not contain the gene and form multicellular structures.
质粒pCB20和pCB22用于在枯草芽孢杆菌细胞中克隆和表达短短芽孢杆菌7882中性蛋白酶基因。基于链球菌质粒pSM19035的复制区域，将13 kb和14 kb含蛋白酶的片段克隆到pCB20质粒中。该基因在枯草芽孢杆菌中表达。应用先前鉴定的表达单元EU19035的营养型启动子可大大提高枯草芽孢杆菌中蛋白酶的表达。表达短短芽孢杆菌中性蛋白酶基因的枯草芽孢杆菌细胞不形成芽孢，比不含该基因的细胞大得多，并形成多细胞结构。
