枯草芽孢杆菌产淀粉酶基因的克隆与表达

Cloning AND Effective Expression Amylase Gene From Bacillus subtilis

本研究以分离自玉溪烤烟叶面的产淀粉酶菌株的基因组DNA为模板,设计特异性引物,通过PCR技术扩增出淀粉酶基因。将目的基因片断与表达载体质粒p ET30a连接,得到重组质粒而后转化到E.coli BL21(DE3),经菌落PCR检测阳性克隆。通过获得重组克隆菌株BL21-p ET,经IPTG诱导表达,SDS-PAGE检测目的蛋白在大肠杆菌中高效表达及获得具有催化活性的重组淀粉酶。

Bacteria strains isolated from the yuxi tobacco leaves surface could effectively produce amylase. Amylase gene from Tobacco leaf surface to produce amylase genome was gained by PCR and cloned into expression vetor p ET30 a.The right plasmid was transformed into E.coli BL21（DE3）,induced expression by IPTG and SDS-PAGE electrophoresis to detected recombinant protein situation in E.coli. Amylase from agents isolated from tobacco leaf surface expressed in E.coli,showed amylase activity.