LfcinB突变基因在大肠杆菌中的融合表达及其活性测定  

LfcinB Mutant Gene Cloned in Escherichia coli Fusion Expression and Activity Determination

作　　者：吴建军[1] 韩跃武[2] 刘凯[1] 张延英[1] 刘永琦[1] 

机构地区：[1]甘肃中医学院,兰州730000 [2]兰州大学基础医学院,兰州730000

出　　处：《生物技术通报》2010年第3期85-89,共5页Biotechnology Bulletin

基　　金：甘肃省科技攻关项目(2GS064-A43-020-21)

摘　　要：测定LfcinB突变基因(mLfcinB)在大肠杆菌中的融合表达及其表达产物的抗菌活性。对LfcinB基因突变后进行克隆并测定序列,mLfcinB基因融合表达载体构建后并在大肠杆菌中的表达,分离提纯表达物并测定其活性。结果显示,Lf-cinB突变基因克隆后表达产物在经SDS-PAGE电泳,考马斯亮蓝G250染色可在29kD处看到预期的融合蛋白条带;对照组空质粒pGEX-4T-1经相同条件诱导后表达的蛋白为26kD。通过抑菌试验发现其对金黄色葡萄球菌ATCC25938具有较强抗菌活性。表明mLfcinB基因表达物具有较强的抗菌作用。The research was to detect LfcinB mutant gene（mLfcinB） in Escherichia coli fusion expression and antibacterial activity of expression products. After LfcinB cloned gene mutations and detection genes sequence, mLfcinB fusion gone expression vector in E. coli after expression,the expression of separate and refine and determine its activity. Results showed that indicated that LfeinB mutant gene cloned in the expression product by SDS-PAGE electrophoresis and Coomassie brilliant blue G250 staining can be seen at the expected fusion 29 kD protein bands,comparing with control group,where the empty plasmid pGEX-4T-1 expression induced by the same conditions, expressed the protein 26 kD. Through the antibacterial test was found on Staphylococcus aureus ATCC25938 has strong antimicrobial activity. Therefore,it proved that mLfcinB complex gone expression has a strong antibacterial effect.

关 键 词：LfcinB基因 突变PCR 融合表达 抗菌 

分 类 号：Q78[生物学—分子生物学]