猪繁殖与呼吸综合征病毒（PRRSV）是一种引起母猪繁殖障碍和仔猪呼吸道症状并导致高死亡率的病毒，给养猪 业带来巨大的损失。2b蛋白是PRRSV的一种小结构蛋白，在病毒的感染和复制中起到关键作用。研究2b蛋白与宿 主细胞蛋白的相互作用，对全面了解PRRSV感染机制与深入认识2b蛋白功能具有重要意义。本研究首先构建猪肺泡巨噬细胞（PAM）cDNA表达文库，利用酵母双杂交技术筛选文库中与2b蛋白有相互作用的蛋白因子，通过酵母回返、双荧光共定位、免疫共沉淀法以及GST-pull down试验对阳性蛋白因子进行验证，并进一步对鉴定的阳性PAM蛋白实施抗体阻断与siRNA干扰表达，以研究阳性PAM蛋白被抑制后PRRSV在PAM上的感染与增殖情况。这将为揭示2b蛋白在PRRSV感染中所起到的作用提供理论基础。

Porcine reproductive and respiratory syndrome virus(PRRSV), cause the severe reproductive problems in sows and high mortality in piglets, and tremendously economic losses in swine breeding industry. 2b protein is a small structural protein of PRRSV, which plays an important role during infection and replication of PRRSV.It is important for fully understanding the infection mechanism of PRRSV and figuring out the functions of 2b protein to study on the interaction of 2b protein and the host cell protein.In this study.We will firstly construct the cDNA expression library of porcine alveolar macrophages (PAM), then Screen the proteins interacting with 2b protein from the cDNA library by yeast two-hybrid system. To verify the positive proteins by yeast turning test,Translocation Fluorescence Cross-correlation tool,Co-Immunoprecipitation and GST-pull down, the antibody blocking and siRNA interference expression experiment will be carried out for the identified positive proteins, that can study the infection and multiplication of PRRSV in the porcine PAM after the inhibition of positive PAM proteins.It should be a helpful rationale for revealing the effect of 2b protein in the process of PRRSV infection.