水稻OsVDAC4诱饵蛋白载体的构建及鉴定  

Construction and Identification of OsVDAC4 Bait Protein Vector in Rice

作　　者：白雪琪 谭艳平[1] 覃永华[1] 王亚楠 杨武 龚秋涵 刘学群[1] 王春台[1] BAI Xue-qi;TAN Yan-ping;QIN Yong-hua(Key Lab for Biotechnology of State Ethnic Affairs Commission,Hubei Provincial Key Laboratory for Protection and Application of Special Plants in Wuling Area of China,College of Life Science,South-Central University for Nationalities,Wuhan,Hubei 430074)

机构地区：[1]中南民族大学生命科学学院,生物技术国家民委重点实验室/武陵山区特色资源植物种质保护与利用湖北省重点实验室,湖北武汉430074

出　　处：《安徽农业科学》2018年第36期90-94,124,共6页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金项目(31170226)

摘　　要：[目的]获得可用于筛选膜蛋白酵母双杂交文库的诱饵蛋白载体。[方法]在对Os VDAC4进行生物信息学分析的基础上,将Os VDAC4全长ORF分别与p BT3-N和p BT3-SUC连接,构建2个诱饵蛋白载体p BT3-Os VDAC4-N-Cub和p BT3-SUC-Os VDAC4-C-Cub,利用膜蛋白酵母双杂交技术鉴定出可用于筛库的诱饵蛋白载体。[结果]p BT3-Os VDAC4-N-Cub和p BT3-SUC-Os VDAC4-C-Cub都可以与目标蛋白互作,但由于Os VDAC4的N端有约50个可自由伸展的氨基酸残基导致p BT3-Os VDAC4-N-Cub本底反应非常强,Os VDAC4的C端存在于膜中而没有可自由伸展的氨基酸残基,p BT3-SUC-Os VDAC4-C-Cub的本底反应很弱。[结论]p BT3-SUC-Os VDAC4-CCub可用于文库筛选,这为获取Os VDAC4的互作蛋白提供了基础。[Objective]To achieve the bait protein vector that suited for screening of the membrane protein yeast two-hybrid library. [Method]On the basis of bioinformatics analysis of Os VDAC4,2 bait protein vectors,p BT3-Os VDAC4-N-Cub and p BT3-SUC-Os VDAC4-C-Cub,were constructed through connecting the full length ORF with p BT3-N and p BT3-SUC respectively. The bait protein vector best suited to screen the library was identified with membrane protein yeast two-hybrid technique. [Result]There were interactions between both p BT3-Os VDAC4-N-Cub and p BT3-SUC-Os VDAC4-C-Cub with the target protein. However,because of about 50 free-stretching amino acid residues in the N-terminal of Os VDAC4,there was a very strong background reaction between p BT3-Os VDAC4-N-Cub and other proteins. And the C-terminal of Os VDAC4 existed in the membrane without free-stretching amino acid residues,p BT3-SUC-Os VDAC4-C-Cub’s background reaction was very weak.[Conclusion]p BT3-SUC-Os VDAC4-C-Cub can be used for library screening,which provides a basis for acquiring proteins interacted with OsVDAC4.

关 键 词：OsVDAC4 诱饵蛋白载体 膜蛋白酵母双杂交 

分 类 号：S188[农业科学—农业基础科学]