芪合酶基因的克隆及其酵母表达载体的构建  

Cloning of Stilbene Synthase Gene and Construction of Its Yeast Expression Vector

作　　者：郭春叶[1] 刘林丽[2] 龚月生[1] 刘勇[1] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]西北农林科技大学生命科学学院,陕西杨凌712100

出　　处：《西北农业学报》2007年第4期67-70,共4页Acta Agriculturae Boreali-occidentalis Sinica

摘　　要：应用基因工程技术将从中国野生属葡萄种高抗白粉病的华东葡萄白河-35-1中分离出的芪合酶基因cDNA片段(Stilbene synthase,STS)进行PCR扩增;把回收的目的基因PCR产物定向插入克隆载体pGEM-TEasy中进行测序与序列分析;采用定向克隆的方法将芪合酶基因克隆到大肠杆菌-酵母菌穿梭型酿酒酵母表达载体pYES6/CT上,转化大肠杆菌,提取阳性重组质粒转化酿酒酵母菌(Saccharomyces cerevisiae)菌株INVSc1,酵母菌落PCR电泳结果显示,在约1 200 bp处有一条亮带,证明芪合酶基因酵母表达载体构建成功,为后期工作的开展奠定了基础,并为开发一种含有白藜芦醇生物活性成分的酵母微生态制剂提供了重要的实验素材。A cDNA fragment of STS gene derived from Vitis pesudoreticulata Baihe35-1 was amplified by PCR. And then the recycling target gene was inserted into the clone vector pGEM-T Easy and carried on sequence analysis. The cloned STS gene was inserted with the right direction into pYES6/CT, a shuttle type plasmid vector （E. coli and S. cerevisiae）, and transformed E. coli. The positive recom- binant vector was selected and transferred into INVScl. The colony PCR showed a band at 1 200 bp and identified the successful construction of yeast expression vector. It may lay a foundation for the later study and provide important experiment materials for developing a yeast micro-ecological agent containing resveratrol.

关 键 词：芪合酶 白藜芦醇 酵母表达载体 构建 微生态制剂 

分 类 号：S816[农业科学—饲料科学] Q943.2[农业科学—畜牧学]