Institute of Preventive Veterinary Science and Key Laboratory of Animal Virology of Ministry of Agriculture, Department of Veterinary Medicine, Zhejiang University, Hangzhou, 310058, China.
Institute of Preventive Veterinary Science and Key Laboratory of Animal Virology of Ministry of Agriculture, Department of Veterinary Medicine, Zhejiang University, Hangzhou, 310058, China.
Virology. 2021 Apr;556:1-8. doi: 10.1016/j.virol.2021.01.011. Epub 2021 Jan 23.
Porcine deltacoronavirus (PDCoV) is one of the emerged coronaviruses posing a significant threat to the swine industry. Previous work showed the presence of a viral accessory protein NS6 in PDCoV-infected cells. In this study, we detected the expression of the NS6 protein in small intestinal tissues of PDCoV-infected piglets. In addition, SDS-PAGE and Western blot analysis of sucrose gradient-purified virions showed the presence of a 13-kDa NS6 protein. Further evidences of the presence of NS6 in the PDCoV virions were obtained by immunogold staining of purified virions with anti-NS6 antiserum, and by immunoprecipitation of NS6 from purified virions. Finally, the anti-NS6 antibody was not able to neutralize PDCoV in cultured cells. These data establish for the first time that the accessory protein NS6 is expressed during infection in vivo and incorporated into PDCoV virions.
猪德尔塔冠状病毒(PDCoV)是一种新兴的冠状病毒,对养猪业构成重大威胁。先前的工作表明,PDCoV 感染细胞中存在一种病毒辅助蛋白 NS6。在本研究中,我们检测到 PDCoV 感染仔猪小肠组织中 NS6 蛋白的表达。此外,通过蔗糖梯度纯化病毒粒子的 SDS-PAGE 和 Western blot 分析显示存在 13kDa 的 NS6 蛋白。通过用抗 NS6 抗血清对纯化病毒粒子进行免疫金染色,以及通过从纯化病毒粒子中免疫沉淀 NS6,获得了 NS6 存在于 PDCoV 病毒粒子中的进一步证据。最后,抗 NS6 抗体不能中和细胞培养中的 PDCoV。这些数据首次确立了辅助蛋白 NS6 在体内感染过程中表达并整合到 PDCoV 病毒粒子中。