一种酿酒酵母组成型分泌表达载体的构建及应用  

The Construction and Application of a Constitutive Secretory Expression Vector of Saccharomyces cerevisiae

作　　者：王国平 欧阳蓓 胡紫艳 刘启铃 赵喜华[1] WANG Guoping;OUYANG Bei;HU Ziyan;LIU Qilin;ZHAO Xihua(College of Life Science,Jiangxi Normal University,Nanchang Jiangxi 330022,China)

机构地区：[1]江西师范大学生命科学学院,江西南昌330022

出　　处：《江西师范大学学报(自然科学版)》2024年第5期509-513,共5页Journal of Jiangxi Normal University(Natural Science Edition)

基　　金：国家自然科学基金(22068014,21666010)资助项目.

摘　　要：该文利用重叠延伸PCR和无缝分子克隆技术构建酿酒酵母组成型分泌表达载体pYAT21,该载体由酿酒酵母组成型分泌表达组件(翻译延伸因子1组成型启动子、α因子N-末端分泌信号肽DNA序列和乙醇脱氢酶2终止子所组成)、2μ复制子、大肠杆菌复制元件以及URA3筛选标记所组成.将草酸青霉16的β-葡萄糖苷酶与pYAT21连接并转化到酿酒酵母中,其发酵上清液粗酶活达到106.8 U·mL^(-1).酿酒酵母组成型分泌表达载体pYAT21可应用于异源蛋白的组成型分泌表达.The recombinant secreted expression vector pYAT21 of S.cerevisiae is constructed by overlapping exten-ded PCR and seamless cloning.pYAT21 vector is constructed,which is composed of a secreted expression component of S.cerevisiae(translation extension factor 1 promoter,αfactor N-terminal secreted signal peptide DNA sequence and alcohol dehydrogenase 2 terminator),2μreplicons,Escherichia coli replication elements,and URA3 screening marker.Theβ-glucosidase of Penicillium oxalicum 16 is linked with pYAT21 and transformed into S.cerevisiae.The crude enzyme activity of the supernatant reaches 106.8 U·mL^(-1).The constitutive secretory expression vector pYAT21 of S.cerevisiae is successfully constructed,which can be used for the constitutive secretory expression of heterologous protein.

关 键 词：酿酒酵母 组成型分泌表达载体 异源蛋白 

分 类 号：Q782[生物学—分子生物学]