摘要
为了探究宿主蛋白2′,5′-寡腺苷酸合成酶2(OAS2)与非洲猪瘟病毒(ASFV)复制之间的关系,本研究通过蛋白免疫印迹(Western-blot)和实时荧光定量PCR(RT-qPCR)分析了ASFV感染对宿主OAS2的调控作用;构建并合成OAS2真核表达质粒以及siRNA干扰序列,通过RT-qPCR、Western-blot方法探究在MA-104细胞中外源过表达OAS2及在PAMs细胞中下调OAS2表达对ASFV体外复制的影响。结果显示,ASFV感染PAMs细胞后,OAS2的蛋白水平和转录水平皆上调,并且外源过表达OAS2显著抑制了ASFV的复制,下调OAS2表达促进了ASFV的复制。本研究通过对宿主蛋白OAS2进行过表达和敲低,证实了OAS2可以抑制ASFV体外复制,该结果为进一步研究宿主对ASFV的抗病毒免疫研究提供了理论基础。
In order to investigate the relationship between host protein 2′,5′-oligoadenylate synthetase 2(OAS2)and the replication of African swine fever virus(ASFV),the present study was conducted to analyze the regulation of OAS2 in the host by ASFV infection by protein immunoblotting(Western-blot)and real-time fluorescence quantitative PCR(RT-qPCR).OAS2 was constructed and synthesized as a eukaryotic expression plasmid and siRNA interfering sequences,and its role was investigated in MA-104 by RT-qPCR and Western-blot.In this study,we analyzed the regulatory effect of ASFV infection on host OAS2 by proteinimmunoblotting(Western-blot)andreal-timefluorescencequantitativePCR(RT-qPCR).Weconstructed and synthesized the eukaryotic expression plasmid of OAS2 and the siRNA interfering sequences,and we investigated the effects of ectopic overexpression of OAS2 in MA-104 cells and the down-regulation of OAS2 in PAMs on the in vitro replication of ASFV by RT-qPCR and Western-blot.The results showed that the protein and transcript levels of OAS2 were up-regulated after ASFV infection in PAMs cells.Moreover,exogenous overexpression of OAS2 significantly inhibited ASFV replication,and down-regulation of OAS2 expression promoted ASFV replication.In this study,we confirmed that OAS2 could inhibit ASFV replication in vitro by overexpressing and knocking down the host protein OAS2,and the results provide a theoretical basis for further research on host antiviral immunity against ASFV.
作者
赵思越
史喜绢
杨行
张大俊
赵登率
陈国辉
陈玲玲
闫文倩
别鑫恬
赵美玉
李平
郑海学
张克山
郜原
ZHAO Siyue;SHI Xijuan;YANG Xing;ZHANG Dajun;ZHAO Dengshuai;CHEN Guohui;CHEN Lingling;YAN Wenqian;BIE Xintian;ZHAO Meiyu;LI Ping;ZHENG Haixue;ZHANG Keshan;GAO Yuan(College of Life Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;State Key Laboratory for Animal Disease Control and Prevention/College of Veterinary Medicine of Lanzhou University/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730000,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2024年第5期577-583,共7页
Chinese Veterinary Science
基金
国家重点研发计划项目(2021YFD1801300)
国家生猪产业体系项目(CARS-35)
中央高校基本科研业务费专项资金资助
国家生猪技术创新中心先导科技项目(NCTIP-XD/C03)
甘肃省科技重大专项(22ZD6NA001)
“十四五”广东省农业科技创新十大主攻方向“揭榜挂帅”项目(2022SDZG02)
中国农业科学院科技创新工程项目(CAAS-ASTIP-2022-LVRI)
中国农业科学院基础科学中心重大任务(CAAS-CSLPDC-202302)。
关键词
非洲猪瘟病毒
OAS2
天然免疫
抑制
African swine fever virus
OAS2
innate immunity
inhibitory
