Biedermann K, Fiedler H, Larsen B S, Riise E, Emborg C, Jepsen P K
Department of Biotechnology, Technical University of Denmark, Lyngby.
Appl Environ Microbiol. 1990 Jun;56(6):1833-8. doi: 10.1128/aem.56.6.1833-1838.1990.
The secretion of a Serratia marcescens nuclease was followed by fermentation with Escherichia coli. A plasmid, p403-SD2, carrying a 1.3-kilobase-pair insert with a 0.4-kilobase-pair region upstream of the nuclease gene caused a growth-phase-regulated expression of nuclease in E. coli in the same way as that seen in S. marcescens. Deletion of the regulatory gene generating plasmid p403-Rsa1 resulted in a constitutive expression of the nuclease. Anaerobiosis stimulated the expression from p403-SD2 in stationary growth phase by a factor of 10 compared with expression stimulated by cultivation in aerobic conditions; no such effect was found for plasmid p403-Rsa1. Different nutritional factors caused the expression level and the amount of extracellular nuclease to vary more when nuclease was expressed from plasmid p403-SD2 than when it was expressed from plasmid p403-Rsa1. A correlation between the regulatory gene and the extracellular secretion of nuclease is proposed.
用大肠杆菌发酵来追踪粘质沙雷氏菌核酸酶的分泌情况。携带一个1.3千碱基对插入片段且在核酸酶基因上游有一个0.4千碱基对区域的质粒p403-SD2,导致大肠杆菌中核酸酶的表达呈现生长阶段调控,其方式与在粘质沙雷氏菌中所见相同。产生质粒p403-Rsa1的调控基因缺失导致核酸酶组成型表达。与在有氧条件下培养所刺激的表达相比,厌氧状态使处于稳定生长阶段的p403-SD2的表达提高了10倍;对于质粒p403-Rsa1未发现这种效应。当从质粒p403-SD2表达核酸酶时,不同的营养因素导致表达水平和细胞外核酸酶量的变化比从质粒p403-Rsa1表达时更大。提出了调控基因与核酸酶细胞外分泌之间的相关性。