基于双向电泳技术的里氏木霉高效组成型启动子筛选  

Screening of constitutively effective promoters in Trichoderma reesei through two dimensional gel electrophoresis

作　　者：禹建腾 谢钧[1] 王绍文[1] 王娟[1] 刘刚[1] 

机构地区：[1]深圳大学生命科学学院深圳市微生物基因工程重点实验室,广东深圳518000

出　　处：《菌物学报》2014年第6期1263-1271,共9页Mycosystema

基　　金：深圳市基础研究计划(No.JCYJ20120613115323982)

摘　　要：获得强启动子是建立高效表达系统的基础。通过蛋白质双向电泳技术从蛋白质水平全面筛选里氏木霉的组成型启动子,获得了3个表达效率较高的启动子,分别是葡萄糖/核糖醇脱氢酶基因的启动子(grdh)、微体蛋白基因的启动子(hex1)和FAD相关蛋白基因的启动子(flo)。通过木聚糖酶Ⅱ(XYN2)的高效同源表达对这些启动子的功能进行了验证,为在里氏木霉中进行同源或异源蛋白的组成型表达提供了有效的工具。Obtaining highly effective promoter is the basis for construction of effective expression system. In this work, two-dimensional gel electrophoresis technology was applied to screen the promoters of highly expressed proteins in the filamentous fungus Trichoderma reesei which was cultivated in glucose containing medium, and three constitutive pro- moters were obtained. They are the promoters of the protein GRDH （glucose/ribitol dehydrogenase）, HEXl （the major component of the Woronin body） and 1FLO （a FAD linked oxidase）, respectively. The effectiveness of these promo- ters was verified through constructing expression system with the gene of XylanaseⅡ （xyn2） as the reporter gene in T. reesei. As a result, all the three promoters effectively conducted the expression of xyn2 in T. reesei in glucose con- taining medium. This work provided effective promoters for expression of recombinant proteins in T. reesei.

关 键 词：里氏木霉 蛋白质组学 二维电泳 木聚糖酶 

分 类 号：Q78[生物学—分子生物学]