禽呼肠孤病毒σC蛋白的真核表达及亚细胞定位的初步分析

Expression of avian reovirus σC protein and subcellular localization

本研究根据GenBenk中σC的序列设计特异性引物,通过PCR扩增σC基因,构建真核表达载体pEF1α-HA-σC,转染DF1细胞,在DF1细胞中表达ARVσC蛋白。结果显示,成功在DF1细胞中表达ARVσC蛋白,大小约为37 ku,Western-blot分析表明,表达的蛋白具有良好的反应原性,激光共聚焦显微镜观察发现在DF1细胞中表达的σC蛋白定位于细胞质中。本研究为今后深入研究σC蛋白在ARV感染过程中的功能和在ARV天然免疫中的具体调控机制奠定了基础。

In this study,we designed specific primers based on the sequence of σC in GenB enk,amplified σC gene by PCR,constructed the eukaryotic expression vector pEF1α-HA-σC,and transfected it into DF1 cells to express ARV σC protein.The results showed that:we successfully expressed the ARV σC protein in DF1 cells with a size of about 37 ku.Western-blot analysis showed that the expressed protein had good reactogenicity.And it was revealed that the expression of σC protein in DF1 cells was localized in the cytoplasm by laser confocal microscopy.This research lays the foundation for the in-depth study of the function of σC protein in the process of ARV infection and its specific regulatory mechanism in natural immunity.