痘苗病毒感染后，宿主细胞的dsRNA依赖的蛋白激酶（PKR）会发生磷酸化，引起细胞微环境改变来对抗病毒；相反，病毒的宿主范围因子（Hrf）K3L蛋白又会调节细胞的PKR抗病毒通路，为病毒的增殖提供适宜的细胞微环境。羊痘病毒是重要的动物痘病毒之一，分析显示羊痘病毒基因组中的K3L同源基因是羊痘病毒的Hrf，但其调节细胞PKR抗病毒信号通路的机制尚不清楚。本课题通过同源重组构建绵羊痘病毒（SPV）K3L基因缺失突变病毒，用细胞培养条件下稳定同位素标记技术（SILAC）结合液质联用质谱（MS/MS）技术鉴定K3L缺失SPV与其亲本SPV感染细胞后，宿主细胞的磷酸化水平差异蛋白和信号通路变化；结合检测突变株对IFNs诱导的PKR途径中关键蛋白磷酸化水平的影响，确定SPV K3L蛋白在调节宿主PKR信号通路中的作用。本课题对解理解宿主与羊痘病毒之间的抗病毒和抗病毒抑制关系以及新的疫苗研制有重要意义。

The host cell proteome and cell micro environment were changed to fight virus, after host cells infected with vaccinia virus. Inversely,virus host range factor(Hrf)can also provide appropriate cell micro environment for virus replication by regulating cells PKR antiviral pathways.However,little is known about the host cell proteome after cells were infected with sheeppox virus(SPV).Bioinformatics showed that vaccinia virus K3L homo-gene in SPV genome are identified as one of the Hrfs in SPV, but its machanisms in rgulating antiviral remian to clarified.A vaccinia K3L gene defective SPV strain(SPVΔK3L)will be constructed through homologous recombination.Then we will detect the host cell phosphorylation level of proteins and signaling pathways by using stable isotope labeling by amino acids in cell culture(SILAC) coupled to liquid MS analysis mass spectrometry(MS/MS) techniques after infection of sheep testicular cells with SPV and SPVΔK3L.At the same time,we will examine the effects of the phosphorylation level in key proteins induced by IFNs in PKR pathways. The roles of sheeppox virus K3L will be identified in regulating the host interferon signal pathway through the above experiments.Our study is of the significance to understand pathogenic mechanism of Capripox virus, and to explain the relations of antiviral response and antiviral inhibition between the host and the SPV as well as develop a novel vaccine for Capripox.