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流感病毒的核衣壳蛋白(NP)是病毒粒子中重要的结构蛋白,参与流感病毒生命周期的多个过程。为筛选与流感病毒NP相互作用的宿主蛋白,本研究应用酵母双杂交技术(Y2H)技术从A549细胞cDNA文库中筛选到与其相互作用的宿主蛋白Tribble 2(TRIB2),并通过免疫共沉淀技术(Co-IP)试验和GST pull down试验进一步证实NP与TRIB2之间存在特异性的直接相互作用。此外,利用激光共聚焦试验证实NP与TRIB2共定位于A549细胞的细胞质内。在A549细胞中过表达TRIB2蛋白后,能够降低流感病毒的滴度,表明TRIB2具有抑制流感病毒的复制功能。本实验为研究流感病毒的复制机理奠定了基础,完善了病毒与宿主相互作用网络。
The nucleocapsid protein (NP) of the influenza virus is an important structural protein in the virus particle and participates in multiple processes of the influenza virus’s life cycle. In order to screen host proteins interacting with influenza virus (NP), yeast two-hybrid assay (Y2H) was used to screen Tribble 2 (TRIB2) which interacts with the cDNA library of A549 cells. The co-immunoprecipitation assay Co-IP test and GST pull down test further confirmed the specific direct interaction between NP and TRIB2. In addition, laser confocal test was used to confirm that NP co-localized with TRIB2 in the cytoplasm of A549 cells. The overexpression of TRIB2 protein in A549 cells decreased the titer of influenza virus, indicating that TRIB2 has the ability to inhibit the replication of influenza virus. This experiment laid the foundation for studying the replication mechanism of influenza virus and perfected the virus-host interaction network.