牛流行热病毒（BEFV）是引起牛流行热的病原，其复制机制研究滞后，制约了抗病毒药物的研发。本团队研究发现，病毒α1蛋白与宿主AnxA2蛋白结合，二者编码基因均促进BEFV的复制，且共转染后有显著的叠加效应。本研究拟利用反向遗传技术拯救α1基因缺失的病毒，检测其增殖及病毒粒子形态是否异常，分析α1基因是否是BEFV复制所必需的基因；利用免疫共沉淀技术筛选与α1结合的病毒其他蛋白，并拯救α1基因上相应结合位点变异的病毒，检测其复制能力及病毒形态，发现与α1协同作用的病毒基因；定位α1上与AnxA2结合的位点，检测相应位点变异对病毒复制的影响；高通量筛选并验证参与α1促进病毒复制的其他候选基因，检测并分析α1促进病毒复制的信号通路；本研究拟以α1基因与病毒其他基因及宿主AnxA2基因的关系为切入点，率先发现α1基因功能，揭示其调控病毒复制的分子机制，以期为抗BEFV药物研发提供靶标分子。

Bovine ephemeral fever virus (BEFV) is the causative agent of bovine ephemeral fever, and little is known of viral replication mechanism, which restricts the development of antiviral drugs. Earlier studies from our team have demonstrated that BEFV alpha 1 protein interacted with the host AnxA2 protein, moreover, each of the genes expression promoted the replication of BEFV and there was an obvious synergistic effects on viral replication when alpha 1 and AnxA2 gene were co-transfected. In this study, the alpha 1 gene deleted BEFV strain will be rescued via reverse genetics technology, then the proliferation and morphology of rescued virus particles will be tested to determine whether alpha 1 gene is required for the replication of BEFV. In addition, the viral proteins combining with BEFV alpha 1 protein and their binding sites will be screened and identified by using coimmunoprecipitation, then replication ability and virus morphology of the rescued BEFV with the corresponding binding site mutation of alpha 1 will be detected, and the viral gene that could be synergistic with the alpha 1 gene will be found. Furthermore, the binding sites between alpha 1 protein and AnxA2 protein will be located and mutated, and then the effects on replication of infectious virus strains based on binding site-specific mutagenesis will be analyzed.What's more, other candidate genes involved in the process of alpha 1 promoting virus replication will be screened and identified by high-throughput sequencing, and then signal transduction pathway of alpha 1 promoting viral replication will be confirmed through regulating the expression of candidate genes. Taken together, it will be the first to reveal the function of BEFV alpha 1 and its molecular mechanism of viral infection from the perspective of the relationship between alpha 1 and viral gene or the host AnxA2, which will provide new target molecule for the research and development of antiviral drugs.