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5. 牛源犬新孢子虫NcAMA1基因酵母双杂交诱饵载体构建及鉴定

牛源犬新孢子虫NcAMA1基因酵母双杂交诱饵载体构建及鉴定

来源 :中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会 | 被引量 : 0次 | 上传用户：zfgzfgzfg

【摘 要】

：

[目的]为筛选牛源犬新孢子虫AMA1与虫体互作的靶蛋白,构建NcAMA1基因酵母双杂交诱饵载体pGBKT7-AMA1.[方法]本试验应用RT-PCR技术从虫体总RNA中扩增了去除跨膜螺旋的AMA1基因,定向克隆到酵母双杂交诱饵载体pGBKT7中,将重组诱饵载体转化酵母Y2H,并验证其在酵母细胞中毒性作用和有无自激活现象.[结果]本试验成功构建了诱饵载体pGBKT7-AMA1,并证明其对酵母细胞无毒

【作 者】

：

李积旭 张守发 贾立军 李男礼 于龙政 薛书江 赵鹏 

【机 构】

：

延边大学农学院 动物医学系,吉林,延吉 133002

【出 处】

：

中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会

【发表日期】

：

2015年6期

【关键词】

：

犬新孢子虫 AMA1 酵母双杂交 诱饵载体 

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　　[目的]为筛选牛源犬新孢子虫AMA1与虫体互作的靶蛋白,构建NcAMA1基因酵母双杂交诱饵载体pGBKT7-AMA1.[方法]本试验应用RT-PCR技术从虫体总RNA中扩增了去除跨膜螺旋的AMA1基因,定向克隆到酵母双杂交诱饵载体pGBKT7中,将重组诱饵载体转化酵母Y2H,并验证其在酵母细胞中毒性作用和有无自激活现象.[结果]本试验成功构建了诱饵载体pGBKT7-AMA1,并证明其对酵母细胞无毒性,且对报告基因无自激活.[结论]本试验构建的诱饵载体pGBKT7-AMA1可用于酵母双杂交系统筛选与虫体互作的宿主蛋白.

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