Barnase在大肠杆菌中的分泌表达和诱导条件优化

Secretive Expression and Optimized Inducement of Barnase

从解淀粉芽孢杆菌中PCR分别扩增解淀粉芽孢杆菌核糖核酸酶barnase基因及其抑制剂barstar基因,采用将barnase基因置于barstar基因保护下的克隆策略,以pET-22b(+)质粒为基础,构建大肠杆菌分泌型表达质粒.IPTG诱导表达目的蛋白后将培养基蛋白进行SDS-PAGE分析并从诱导温度、IPTG诱导浓度和诱导时间三方面初步优化诱导表达条件.

Using PCR method to amplify Bacillus amyloliquefaciens ribonuclease barnase gene and its inhibitor barstar gene from Bacillus amyloliquefaciens. The secretive expression vector is constructed by the strategy of protective clone from plasmid pET-22b （＋）. After induced by IPTG, a 12 kD target barnase protein can be detected from medium protein by SDS-PAGE analysis. Inducing conditions of temperature, IPTG concentration and inducing time have also been optimized.