中性蛋白酶基因在大肠杆菌中诱导表达条件的研究  

Research on Expression Condition of Neutral Protease in the E.coli

作　　者：张敏[1,2] 赵丛[1] 路福平[1] 赵洪坤[1] 杜连祥[1] 

机构地区：[1]天津科技大学生物工程学院,天津市工业微生物重点实验室,天津300457 [2]沈阳农业大学工程学院

出　　处：《食品与发酵工业》2007年第10期31-34,共4页Food and Fermentation Industries

摘　　要：采用PCR方法从枯草芽孢杆菌中扩增得到中性蛋白酶基因npr,与表达载体pET-22b(+)连接构建成重组质粒pET22b-npr,转化大肠杆菌BL21得到重组工程菌株。经IPTG诱导,其所含有的中性蛋白酶基因可高效表达。研究不同的表达条件对中性蛋白酶表达水平的影响,发现当培养液的OD_(600)值达到0.6～0.8时,添加诱导剂IPTG至终浓度为0.8 mmol/L,28℃诱导7h,重组工程菌中性蛋白酶的表达量最高,SDS-PAGE电泳结果显示出明显的分子质量约43 ku的特异性蛋白条带。The neutral protease gene from Bacillus subtilis was amplified by PCR and coloned into pET- 22b plasmid to create the recombinant plasmid pET22b-npr. Then we got the recombinant strain by transforming pET22b-npr into E. coli BL21. When it was induced by IPTG, the neutral protease could be produced with a high level. In this study, the optimal condition of expression was obtained. When the OD600 of culture reached 0.6-0. 8, IPTG was added to give a final concentration of 0. 8 mmol/L. Incubating the culture at 28℃ for 7h, the expression content of neutral protease reached the most. The specific protein band about 43 ku was shown in the SDS PAGE gel.

关 键 词：枯草芽孢杆菌 大肠杆菌 中性蛋白酶 诱导表达 

分 类 号：Q78[生物学—分子生物学]