利用酵母单杂交方法筛选大豆根融合基因表达cDNA文库  

Construction of Fusion Gene Expression cDNA Library of Soybean Root with Yeast One-hybrid Method

作　　者：刘清醒[1] 郭长虹[1] 毕影东[2] 郭东林[1] 

机构地区：[1]哈尔滨师范大学生命科学与技术学院/黑龙江省分子细胞遗传与遗传育种重点实验室,黑龙江哈尔滨150025 [2]黑龙江省农业科学院耕作栽培研究所,黑龙江哈尔滨150086

出　　处：《大豆科学》2013年第2期165-167,共3页Soybean Science

基　　金：国家转基因生物新品种培育重大专项(2011ZX08004-002);哈尔滨科技创新人才研究专项资金(RC2011LX002004)

摘　　要：酵母单杂交体系通过筛选cDNA文库可直接得到与靶序列相互作用的蛋白质基因序列,是研究DNA-蛋白质相互作用和转录因子功能的最常用、最有效的方法之一。通过构建与Gal4p的激活结构域融合的表达cDNA文库,利用抗病相关基因启动子区的W-box顺式元件采用酵母单杂交方法对该文库进行了筛选,共得到4个阳性克隆。结果为进一步研究与顺式元件W-box相互作用的WRKY蛋白奠定了基础。Gene sequence of protein which interacted with specific cis-element can be found by screening cDNA library in yeast one-hybrid system. Yeast one-hybrid system has become an important efficient method to investigate DNA-protein interac- tion and function of transcription factor. In this report,gene expression cDNA library fused to activation domain of Gal4p was constructed. Library was investigated using W-box cis-element in yeast one-hybrid system. As a result,4 positive clones were found. The study paved a foundation for screening WRKY protein which interacted with W-box cis-element.

关 键 词：大豆 CDNA文库 酵母单杂交 

分 类 号：S565.1[农业科学—作物学]