穿山龙薯蓣皂苷糖苷酶基因的亚克隆及表达  

Sub-cloning and Expression of Dioscorea nipponica Glycosidase Gene

作　　者：王庆宇[1] 金凤燮[1] 鱼红闪[1] 

机构地区：[1]大连工业大学生物工程学院,辽宁大连116034

出　　处：《安徽农业科学》2011年第10期5799-5802,共4页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金项目(30371744;30470055;20476017);辽宁省科学技术基金项目(20042132);辽宁省教育厅创新团队项目(2009T007)

摘　　要：[目的]克隆并表达穿山龙薯蓣皂苷糖苷酶基因。[方法]将穿山龙薯蓣皂苷糖苷酶基因亚克隆到毕赤酵母表达载体pPIC9K,再将构建后的表达载体电转化到毕赤酵母GS115中,并诱导表达该基因。[结果]重组酵母菌在0.5%甲醇中培养144 h后,所提取的酶蛋白具有穿山龙薯蓣皂苷糖苷酶的活性,经SDS-PAGE分析,确定其相对分子质量为58 kD。[结论]穿山龙薯蓣皂苷糖苷酶基因得到了表达。[Objective]To sub-clone and express glycosidase gene of Dioscorea nipponica.[Method]The glycosidase gene of Dioscorea nipponica was sub-cloned into Pichia pastoris GS115 expression vector pPIC9K,and the constructed expression vector was transformed into Pichia pastoris GS115 by electroporation method to induce the glycosidase gene.[Result]After have been cultivated for 144 hours in 0.5% methanol,the glycosidase gene in recombinant was proved to be active,and the relative molecular weight of which was 58 kDu through SDS-PAGE analysis.[Conclusion]Glycosidase gene of Dioscorea nipponica was expressed successfully.

关 键 词：穿山龙薯蓣皂苷糖苷酶 巴斯德毕赤酵母 基因表达 

分 类 号：S567[农业科学—中草药栽培]