植物甜蛋白des-pGlu1-Brazzein基因在乙醇氧化酶缺陷型甲醇酵母中的表达及其活性分析  

Expression of Plant des-pGIul-Brazzein Gene in Alcohol Oxidasedefective Strain of Pichia pastoris and Analysis of Its Biological Activity

作　　者：徐婷婷[1] 陈锐博[1] 暴元元[1] 赵卫东[1] 郑振宇[1] 李春丽[1] 

机构地区：[1]河南农业大学牧医工程学院,郑州450002

出　　处：《生物技术通报》2012年第12期139-143,共5页Biotechnology Bulletin

摘　　要：采用BglⅡ酶切重组表达载体pPIC9K-Bra,纯化后电击转化甲醇酵母菌GS115,构建乙醇氧化酶缺陷型表达菌株GS115-pPIC9K-Bra,筛选鉴定后,以0.5%的甲醇进行诱导,表达的目的蛋白约占上清总蛋白的95%,纯度较高,并具有一定的甜度。成功构建了乙醇氧化酶缺陷型的甲醇酵母表达菌株,为深入研究其应用奠定了基础。Digested by Bgl Ⅱ, the linear recombinant expression vector pPIC9K-Bra was transformed into Pichia pastoris GSl15 by electric shock to produce alcohol oxidase -defective expression strain GS115-pPIC9K-Bra. The strain was identified by PCR and induced by 0.5% methanol. The resuhs indicated that the secreted target protein accounted for 95% of total protein in supernatant fluid and showed biological activity. The alcohol oxidase -defective expression strain GS115-pPIC9K-Bra was successfully constructed which was foundation for further study.

关 键 词：甜蛋白 甲醇酵母 克隆表达 活性检测 

分 类 号：Q78[生物学—分子生物学]