宿主OAS2蛋白抑制非洲猪瘟病毒体外复制的研究
Inhibition of African swine fever virus replication by the host OAS2 protein in vitro
赵思越 1史喜绢 1杨行 2张大俊 2赵登率 2陈国辉 1陈玲玲 2闫文倩 2别鑫恬 2赵美玉 1李平 2郑海学 2张克山 2郜原3
作者信息
- 1. 甘肃农业大学 生命科学技术学院,甘肃 兰州 730070;中国农业科学院 兰州兽医研究所 兰州大学动物医学与生物安全学院 动物疫病防控全国重点实验室,甘肃 兰州 730000
- 2. 中国农业科学院 兰州兽医研究所 兰州大学动物医学与生物安全学院 动物疫病防控全国重点实验室,甘肃 兰州 730000
- 3. 甘肃农业大学 生命科学技术学院,甘肃 兰州 730070
- 折叠
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.
Abstract
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 protein immunoblotting(Western-blot)and real-time fluorescence quantitative PCR(RT-qPCR).We constructed 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.
关键词
非洲猪瘟病毒/OAS2/天然免疫/抑制Key words
African swine fever virus/OAS2/innate immunity/inhibitory引用本文复制引用
基金项目
国家重点研发计划(2021YFD1801300)
国家生猪产业技术体系建设项目(CARS-35)
中央高校基本科研业务费专项()
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)
广东省农业科技创新十大主攻方向"揭榜挂帅"项目(十四五)(2022SDZG02)
中国农科院科技创新工程项目(CAAS-ASTIP-2022-LVRI)
中国农科院基础科学中心重大任务项目(CAAS-CSLPDC-202302)
出版年
2024
国家科技期刊平台
NSTL
万方数据