新城疫病毒HN蛋白酵母双杂交诱饵载体的构建及鉴定  

Construction and identification of HN bait vector of newcastle disease virus in yeast two-hybrid system

作　　者：胡湘云[1] 高小龙[1] 付向晶[1] 王燕平[1] 刘丹丹[1] 常旭东[1] 刘蓬[1] 杜恩岐[1] 王兴龙[1] 党如意[1] 杨增岐[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2016年第7期45-49,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金项目(31272578)

摘　　要：【目的】应用酵母双杂交技术构建新城疫病毒的血凝素-神经氨酸酶(HN)蛋白线性中和表位区的诱饵载体pGBKT7-HN-neu,为筛选靶向新城疫病毒HN蛋白的中和性纳米抗体奠定基础。【方法】合成HN蛋白的线性中和表位区HN-neu,将其克隆至酵母双杂交系统诱饵载体pGBKT7中,构建诱饵载体pGBKT7-HN-neu,经酶切鉴定和测序验证正确后,将pGBKT7-HN-neu转化酵母菌Y2H Gold,检测其在酵母细胞中的毒性和自激活作用。【结果】成功合成了HN-neu,构建的pGBKT7-HN-neu在酵母细胞Y2H Gold中无毒性和自激活能力。【结论】成功构建了诱饵载体pGBKT7-HN-neu。[Objective] The recombinant bait vector pGBKTT-HN-neu was constructed by yeast two- hybrid system to select variable domain of heavy chain antibody（VHH）against newcastle disease virus （NDV） HN protein. [Method] The neutralizing epitope of NDV HN protein was synthetized and cloned into bait vector pGBKT7 of yeast two-hybrid system. After being verified by enzyme digestion and sequencing, pGBKTT-HN-neu was transformed into yeast cells Y2H Gold. Then, self-activation and toxic action of the bait vector pGBKTT-HN-neu were tested. [Result] HN-neu was successfully synthetized and the recombination bait vector pGBKTT-HN-neu was proven to be no self-activation and not toxic to the host yeast cell. [Conclusion] The bait vector pGBKTT-HN-neu was successfully constructed.

关 键 词：新城疫病毒 HN蛋白 酵母双杂交系统 诱饵载体 

分 类 号：S852.65[农业科学—基础兽医学]