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

生物技术通报 ›› 2012, Vol. 0 ›› Issue (12): 139-143.

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

徐婷婷, 陈锐博, 暴元元, 赵卫东, 郑振宇, 李春丽

河南农业大学牧医工程学院，郑州 450002

收稿日期:2012-05-11 修回日期:2013-01-25 出版日期:2012-12-26 发布日期:2013-02-06
作者简介:徐婷婷, 女, 硕士研究生, 研究方向:基因工程;E-mail:sdxutingting@sian.com

Expression of Plant des-pGlu1-Brazzein Gene in Alcohol OxidasedefectiveStrain of Pichia pastoris and Analysis of Its Biological Activity

Xu Tingting, Chen Ruibo, Bao Yuanyuan, Zhao Weidong, Zheng Zhenyu, Li Chunli

Department of Animal Science and Veterinary Medicine，Henan Agricultural University，Zhengzhou 450002

Received:2012-05-11 Revised:2013-01-25 Published:2012-12-26 Online:2013-02-06

摘要/Abstract

摘要： 采用Bgl Ⅱ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。

关键词: 甜蛋白, 甲醇酵母, 克隆表达, 活性检测

Abstract: Digested by Bgl Ⅱ, the linear recombinant expression vector pPIC9K-Bra was transformed into Pichia pastoris GS115 byelectric shock to produce alcohol oxidase -defective expression strain GS115-pPIC9K-Bra. The strain was identified by PCR and induced by 0.5%methanol. The results indicated that the secreted target protein accounted for 95% of total protein in supernatant fluid and showed biologicalactivity. The alcohol oxidase -defective expression strain GS115-pPIC9K-Bra was successfully constructed which was foundation for furtherstudy.

徐婷婷, 陈锐博, 暴元元, 赵卫东, 郑振宇, 李春丽. 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[J]. 生物技术通报, 2012, 0(12): 139-143.

Xu Tingting, Chen Ruibo, Bao Yuanyuan, Zhao Weidong, Zheng Zhenyu, Li Chunli . Expression of Plant des-pGlu1-Brazzein Gene in Alcohol OxidasedefectiveStrain of Pichia pastoris and Analysis of Its Biological Activity[J]. Biotechnology Bulletin, 2012, 0(12): 139-143.

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