Yang Chee-Hing, Li Hui-Chun, Hung Cheng-Huei, Lo Shih-Yen
Institute of Medical Sciences, Tzu Chi University, Hualien, 97004, Taiwan.
Methods Mol Biol. 2015;1282:197-212. doi: 10.1007/978-1-4939-2438-7_17.
To understand the molecular mechanisms of viral replication and pathogenesis, it is necessary to establish the virus-host protein interaction networks. The yeast two-hybrid system is a powerful proteomic approach to study protein-protein interactions. After the identification of specific cellular factors interacting with the target viral protein using the yeast two-hybrid screening system, co-immunoprecipitation and confocal microscopy analyses are often used to verify the virus-host protein interactions in cells. Identification of the cellular factors required for viral survival or eliminating virus infected cells could help scientists develop more effective antiviral drugs. Here we summarize a standard protocol used in our lab to study the coronavirus-host protein interactions, including yeast two-hybrid screening, co-immunoprecipitation, and immunofluorescence microscopy analyses.
为了解病毒复制和发病机制的分子机制，有必要建立病毒-宿主蛋白相互作用网络。酵母双杂交系统是研究蛋白质-蛋白质相互作用的一种强大的蛋白质组学方法。使用酵母双杂交筛选系统鉴定与目标病毒蛋白相互作用的特定细胞因子后，通常使用免疫共沉淀和共聚焦显微镜分析来验证细胞中的病毒-宿主蛋白相互作用。鉴定病毒存活所需的细胞因子或清除病毒感染细胞有助于科学家开发更有效的抗病毒药物。在这里，我们总结了我们实验室用于研究冠状病毒-宿主蛋白相互作用的标准方案，包括酵母双杂交筛选、免疫共沉淀和免疫荧光显微镜分析。
