为鉴定猪瘟病毒(CSFV)衣壳蛋白(C)与宿主细胞核仁素(NCL)蛋白的相互作用,本研究将重组质粒pEGFP-CSFV-C转染至猪睾丸细胞(ST)中,采用GST pull-down和免疫共沉淀方法沉淀ST细胞中的NCL蛋白,通过共聚焦显微镜分析C蛋白和NCL蛋白的共定位情况.结果表明,CSFV C蛋白与NCL蛋白在核仁中存在共定位现象,并且在体外和转染细胞中均存在相互作用.NCL蛋白被抑制后,CSFV的复制能力下降.本研究为进一步分析C蛋白核转运机制以及在CSFV复制过程中的功能提供了实验依据.
To identify the interaction between classical swine fever virus (CSFV) capsid (C) protein and cellular nucleolin (NCL) in cells, the methods of GST pull-down and Co-IP were used precipitate the NCL protein of swine testicle (ST) cells, respectively. The results showed that cellular NCL interacts with C protein of CSFV in vitro and in transfected cells. Moreover, NCL knock down impair CSFV replication. These results provided the basis for further function studies of C transport and C protein in CSFV replication.