畜牧兽医学报2018，Vol.49Issue(1)：139-146,8.DOI:10.11843/j.issn.0366-6964.2018.01.016
抗病毒蛋白Viperin抑制猪瘟病毒在PK-15细胞上的复制
Porcine Anti-viral Protein Viperin Inhibits the Replication of Classical Swine Fever Virus in PK-15 Cells
摘要
Abstract
Viperin is an antiviral protein which could inhibit the replication of a wide range of viruses.The aim of this study was to explore the anti-CSFV activity of porcine Viperin protein.CSFV was inoculated in the cell line of PK-Vi over-expressing Viperin,viral load was detected by real-time qRT-PCR and virus titration.Knockdown of Viperin expression in PK-Vi by siRNA (siVi) was performed and CSFV replication was detected.Viral load in cell culture supernatants and cell lysates were examined to analyze the influence of Viperin on virus release.The co-localization and interaction of Viperin with CSFV E2 protein was determined by confocal laser scanning microscopy test and co-immunoprecipitation (Co-IP) assay.The genome copy numbers and viral titers of CSFV in PK-Vi was significantly decreased by 68.75％,83.61％,77.27％ and 68.75％,87.5％,80.39％ at 24,48 and 72 hpi (P＜0.05),comparing with control cells (PK-C1 expressing EGFP).Knockdown of Viperin expression retrieved the replication of CSFV,which was significantly higher than those in PK-Vi and siNC transfected PK-Vi (P＜0.05) although it was still lower than those of PK-C1 and siRNA treated PK-C1 groups.Viperin expression had no effect on virus release from PK 15 cells.Confocal laser scanning microscopy test showed Viperin protein coqocalized with E2 protein in CSFV infected cells and plasmid transfected 293T cells.Co-IP assay indicated that Viperin could interact with E2 protein.Porcine Viperin protein effectively inhibited CSFV replication in vitro,potentially via interaction of Viperin with CSFV proteins.The results provide foundation for further studies of the interaction of CSFV infection with host immune response.关键词
Viperin/猪瘟病毒/抗病毒/共定位/相互作用Key words
Viperin/CSFV/inhibition/co-localization/interaction分类
农业科技引用本文复制引用
李文良,毛立,邓加武,郝飞,李基棕,杨蕾蕾,张纹纹,江杰元..抗病毒蛋白Viperin抑制猪瘟病毒在PK-15细胞上的复制[J].畜牧兽医学报,2018,49(1):139-146,8.基金项目
国家自然科学基金(31402180) （31402180）
江苏省自然科学基金(BK20130729) （BK20130729）
