Detection of RNA-dependent RNA polymerase of porcine epidemic diarrhea virus.

Suya Liao, Yijing Chen, Yulan Yang, Guanhua Wang, Qihang Wang, Jiaxin Liu, Haowen Wu, Qiyuan Luo, Yun Chen

Journal of immunological methods 2023 Apr

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The RNA synthesis of porcine epidemic diarrhea virus (PEDV) is a sophisticated process performed by a multilingual viral replication complex, together with cellular factors. A key enzyme of this replication complex is RNA-dependent RNA polymerase (RdRp). However, there is limited knowledge about PEDV RdRp. In our present study, a polyclonal antibody against RdRp was prepared by using a prokaryotic expression vector pET-28a-RdRp to study the function of PEDV RdRp and provide a tool to investigate PEDV pathogenesis. In addition, the enzyme activity and half-life of PEDV RdRp were investigated. The result showed that the polyclonal antibody against PEDV RdRp was successfully prepared and was able to be used to detect PEDV RdRp by immunofluorescence and western blotting. Additionally, enzyme activity of PEDV RdRp reached nearly 2 pmol/μg/h and the half-life of PEDV RdRp was 5.47 h. Copyright © 2023 Elsevier B.V. All rights reserved.

Citation

Suya Liao, Yijing Chen, Yulan Yang, Guanhua Wang, Qihang Wang, Jiaxin Liu, Haowen Wu, Qiyuan Luo, Yun Chen. Detection of RNA-dependent RNA polymerase of porcine epidemic diarrhea virus. Journal of immunological methods. 2023 Apr;515:113442