北京棒杆菌高丝氨酸脱氢酶基因的克隆与表达产物活性测定

摘要

Based on the DNA sequence of Beijing corynebacterium E31, homoserine dehydrogenase gene was amplified. Structural gene was cloned into downstream of T7 promote and expression plasmid was obtained. The expression plasmid was transferred to E.coli BL21 （DE3）and induced by IPTG, and high expression was obtained. The analysis of enzyme activity indicated that homoserine dehydrogenase of recombinant Escherichia coli BL21 was 10 times as control bacteria.%以北京棒杆菌E51染色体DNA为模板，用POP．法扩增了高丝氨酸脱氢酶（HD）基因。将结构基因装载于pET-28a质粒的T7启动子下游，得到了重组表达质粒pET-HD。将其转入大肠杆菌（E．coli）BL21（DE5），经异丙基硫代半乳糖苷（IPTG）诱导，获得了高效的表达。含表达质粒的茵体胞内粗提液经酶活分析表明，高丝氨酸脱氢酶的活性为不含重组质粒的对照茵的10倍。