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.
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2007年第10期31-34,共4页
Food and Fermentation Industries
