3型鲤疱疹病毒白介素10单克隆抗体制备与鉴定OA北大核心CSTPCD

Cyprinid herpesvirus 3(CyHV3),also known as Koi herpesvirus(KHV),is a highly contagious pathogen that threatens the sustainability of cyprinid aquaculture.The ORF134 gene of CyHV3 encodes an interleukin(IL)10 like protein.In this study,we constructed a prokaryotic expression plasmid of ORF134 and transformed it into the E.coli Rosetta(DE3)cells for expression of recombinant protein.The recombinant CyHV3-IL10 protein was purified by size exclusion chromatography and used for immunization of mice to generate monoclonal antibodies.The antibodies were then characterized by ELISA,Western blotting and immunofluorescent assay.SDS-PAGE analysis showed that the recombinant CyHV3-IL10 protein was approximately 18 ku in size and was expressed mainly in the form of inclusion bodies.The inclusion bodies were denatured,refolded and subjected to size exclusion chromatography.The recombinant CyHV3-IL10 protein was of high purity.Western blotting and immunofluorescence analyses revealed that the CyHV3-IL10 monoclonal antibodies recognized the recombinant CyHV3-IL10 protein expressed in the E.coli and HEK293 cells and could specifically detect the HEK293 cells expressing the CyHV3-IL10 protein.In summary,the availability of CyHV3-IL10 monoclonal antibodies lays the foundation for the functional study of CyHV3-IL10 and the establishment of CyHV3 diagnostic tools,and provides novel strategies for the prevention and control of the diseases caused by CyHV3.