Zappe H, Blatch G L, Woods D R
Department of Microbiology, University of Cape Town, South Africa.
J Gen Microbiol. 1992 Feb;138(2):319-27. doi: 10.1099/00221287-138-2-319.
Previously we reported [Deane, S. M., Maharaj, R., Robb, F. T. & Woods, D. R. (1987) Journal of General Microbiology 133, 2295-2302] that the production of a Vibrio alginolyticus SDS-resistant alkaline serine protease (Pro A) cloned in Escherichia coli was characterized by a 12 h delay between the synthesis of an inactive precursor and secretion of active Pro A. Replacement of the V. alginolyticus promoter region by the alpha-amylase promoter region from Bacillus amyloliquefaciens resulted in the simultaneous synthesis and secretion of Pro A in E. coli. The V. alginolyticus pro A gene cloned on a shuttle vector did not produce active Pro A in Bacillus subtilis. Although Pro A has a typical Gram-positive signal sequence, it was not functional in B. subtilis. Replacement of the Pro A signal sequence with the alpha-amylase signal sequence resulted in the production of active Pro A in B. subtilis.
此前我们报道过[迪恩，S.M.，马哈拉杰，R.，罗布，F.T.和伍兹，D.R.（1987年）《普通微生物学杂志》133卷，2295 - 2302页]，克隆于大肠杆菌中的溶藻弧菌抗十二烷基硫酸钠碱性丝氨酸蛋白酶（Pro A）的产生，其特征在于无活性前体的合成与活性Pro A的分泌之间有12小时的延迟。用解淀粉芽孢杆菌的α -淀粉酶启动子区域替换溶藻弧菌的启动子区域，导致Pro A在大肠杆菌中同时合成和分泌。克隆在穿梭载体上的溶藻弧菌pro A基因在枯草芽孢杆菌中不产生活性Pro A。尽管Pro A具有典型的革兰氏阳性信号序列，但它在枯草芽孢杆菌中不起作用。用α -淀粉酶信号序列替换Pro A信号序列，导致在枯草芽孢杆菌中产生活性Pro A。
