Identification of structure proteins of cyprinid herpesvirus 2
2020
Gao, Wa | Wen, Hong | Wang, Hao | Lu, Jiaquan | Lu, Liqun | Jiang, Yousheng
Cyprinid herpesvirus 2 (CyHV-2) is the causative agent of herpesviral hematopoietic necrosis (HVHN). In order to identify the protein contained the genome of CyHV-2, virus particles were purified by sucrose density gradient in combination with ultracentrifugation and observed by transmission electron microscope. The viral proteins were separated by SDS-PAGE, and then identified by mass spectrometry identification. The anti-CyHV-2 polyclonal antibody prepared by immunizing mice with purified virus was subjected to Western blotting analysis and mass spectrometry identification to identify the major immunogenic proteins to mice. A large number of intact CyHV-2 virions were collected from the sucrose layer between 50% and 66%, and a small amount of viral envelops were damaged. The results of mass spectrometry identification showed that CyHV-2 containing 74 proteins, including 3 capsid proteins, 18 membrane proteins, 53 other proteins; Western blotting and mass spectrometry identification showed that there are eight major immunogenic proteins, namely pORF92, pORF115, pORF25, pORF57, pORF66, pORF72, pORF131 and pORF132. This study is the first to identify the proteins contained in CyHV-2 as well as the major immunogenic proteins, which will provide a more comprehensive theoretical and practical basis for the in-depth study of CyHV -2 virus.
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