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

作　　者：单妍 王毅 吴丽君 马永凯 宋鹏飞 魏云林[3] Shan Yan;Wang Yi;Wu Li-jun;Ma Yong-kai;Song Peng-fei;Wei Yun-lin(Haiyuan CollegeKunming Medical University,YunnanKunming 650106;China Tobacco Yunnan IndustrialCo.,Ltd.,YunnanKunming 653100;Biotechnology Research Center,Kunming University of Science and Technology,YunnanKunming 650500)

机构地区：[1]昆明医科大学海源学院,云南昆明650106 [2]云南中烟工业有限责任公司,云南昆明653100 [3]昆明理工大学生物工程技术研究中心,云南昆明650500

出　　处：《生物化工》2018年第3期4-7,共4页Biological Chemical Engineering

基　　金：云南中烟有限责任公司科技开发项目(2016YL03;S-6013067)

摘　　要：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.

关 键 词：Α-淀粉酶 枯草芽孢杆菌 克隆 表达 

分 类 号：Q785[生物学—分子生物学]