中国动物传染病学报2016，Vol.24Issue(1)：62-69,8.
柔嫩艾美耳球虫钙依赖蛋白激酶4酵母双杂交诱饵质粒的构建与检测
DETECTION AND CONSTRUCTION OF EIMERIATENELLA CDPK4 BAIT PLASMID FOR YEAST TWO-HYBRID SYSTEM
摘要
Abstract
In order to screen for the interacting protein with CDPK4 of Eimeriatenella, the recombinant bait plasmid pGBKT7-EtCDPK4 was constructed, which can be used to screen for Eimeriatenella yeast two-hybrid cDNA library. The total RNA of E.tenellasporozoite was extracted, then the first strand of cDNA was obtained by reverse transcriptase. The 1660 bp fragment of EtCDPK4 was amplified by PCR and cloned into yeast BD bait vector pGBKT7, then the recombinant plasmid pGBKT7-EtCDPK4 was identified by double enzyme digestion and sequenced. The self-activation, toxic action and protein expression of the recombination bait plasmid pGBKT7-EtCDPK4 were tested, respectively. These results show that the recombinant pGBKT7-EtCDPK4 plasmid was constructed successfully and proved to be no self-activation and toxic action. Moreover, it was expressed in the Y2HGold yeast cell. So the recombinant plasmid pGBKT7-EtCDPK4 can be used to screen for Eimeriatenella sporozoites yeast two-hybrid cDNA library.关键词
柔嫩艾美耳球虫/钙依赖蛋白激酶/诱饵质粒/自激活作用/细胞毒性Key words
Eimeriatenella/CDPK/bait plasmid/self-activation/toxic effect分类
农业科技引用本文复制引用
王自文,黄兵,董辉,赵其平,夏伟丽,朱顺海,门启婓,李聪,朱雪龙,韩红玉..柔嫩艾美耳球虫钙依赖蛋白激酶4酵母双杂交诱饵质粒的构建与检测[J].中国动物传染病学报,2016,24(1):62-69,8.基金项目
国家自然科学基金（31572266）；中央级公益性科研院所基本科研业务费专项资金项目 （）
