| 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。 |
| 关键词: 新城疫病毒 HN蛋白 酵母双杂交系统 诱饵载体 |
| DOI： |
| 分类号: |
| 基金项目:国家自然科学基金项目（31272578） |
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| Construction and identification of HN bait vector of newcastle disease virus in yeast two-hybrid system |
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HU Xiangyun,GAO Xiaolong,FU Xiangjing,et al
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| Abstract: |
| 【Objective】The recombinant bait vector pGBKT7-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,pGBKT7-HN neu was transformed into yeast cells Y2H-Gold.Then,self-activation and toxic action of the bait vector pGBKT7-HN-neu were tested. 【Result】HN neu was successfully synthetized and the recombination bait vector pGBKT7-HN-neu was proven to be no self-activation and not toxic to the host yeast cell.【Conclusion】The bait vector pGBKT7-HN-neu was successfully constructed. |
| Key words: newcastle disease virus (NDV) hemagglutinin-neuraminidase (HN) yeast two-hybrid system bait vector |
