GOPC对猪瘟病毒在PK-15细胞中增殖的影响  

Effect of GOPC Knockdown on CSFV Proliferation in a PK-15 Cell Line

作　　者：何俊瑶 宋梦昭 纪甜甜 罗燕 邓文[1] HE Jun-yao;SONG Meng-zhao;JI Tian-tian;LUO Yan;DENG Wen(College of Veterinary Medicine,Northwest A&F University,Yangling Shaanxi,712100,China)

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100

出　　处：《动物医学进展》2025年第3期1-5,共5页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(32272998)。

摘　　要：旨在探究高尔基体支架蛋白GOPC对猪瘟病毒(CSFV)增殖的影响,为其在CSFV增殖过程中的作用和分子机制研究奠定基础。设计合成3条靶向猪GOPC基因的siRNA,验证对GOPC mRNA的干扰效率。以高效siRNA序列为基础,干扰GOPC在PK-15细胞表达,接毒后检测GOPC敲低情况下CSFV的增殖状况。同时用RT-qPCR和Western blot分析CSFV感染后PK-15细胞中GOPC的表达情况。结果表明,GOPC敲低的PK-15细胞中CSFV基因组拷贝数显著上升,IFA也检测到GOPC敲低后CSFV E2蛋白表达量显著增高。在CSFV感染PK-15细胞后,随着病毒感染时间的延长,宿主细胞GOPC的表达量出现了下调趋势,但在感染后24 h出现短暂上调。因此,从上述结果可以初步推断GOPC蛋白抑制CSFV的增殖复制,而CSFV则可以通过调节细胞GOPC蛋白表达从而促进自身增殖。By investigating the effect of GOPC knockdown on CSFV proliferation,this study aims to lay a foundation for the molecular mechanisms of CSFV proliferation and offer new insight for prevention of the CSFV infection.Firstly,three siRNAs were designed and synthesized to validate their interference efficiency on knockdown of GOPC.Subsequently,GOPC was knocked-down in PK-15 cell,followed by virus inoculation to assess the impacts of GOPC knockdown on CSFV proliferation.Meanwhile,we utilized RT-qPCR and Western blot assays to analyze the expression of GOPC in PK-15 cells after CSFV infection.The result indicated a significant increase of CSFV genome copy numbers after GOPC knockdown,and IFA assay also demonstrated a significant increase of CSFV E2 protein expression levels following interference expression of GOPC protein.In addition,the impact of CSFV infection on GOPC expression was further examined in PK-15 cells,showing varied levels of GOPC protein expression at different time points post-CSFV infection.These results suggest that GOPC protein inhibits CSFV proliferation in host cells and CSFV regulates GOPC protein to promote its proliferation.

关 键 词：猪瘟病毒 GOPC蛋白 病毒增殖 

分 类 号：S852.651[农业科学—基础兽医学]