Identification of a novel envelope protein encoded by ORF 136 from Cyprinid herpesvirus 3

Abstract

Cyprinid herpesvirus 3 (CyHV-3) is the pathogenic agent of koi herpesvirus disease (KHVD) afflicting common carp and koi (Cyprinus carpio L.) populations globally. As described previously, proteomic analyses of purified CyHV-3 particles have shown that at least 46 structural proteins are incorporated into CyHV-3 virions; among these ORF136 may encode a putative envelope protein. In this study, Western blotting analysis showed that a specific band with the predicted molecular weight of 17 kDa was detected both in purified virions and envelope components using a rabbit anti-ORF136 polyclonal antibody. Indirect immunofluorescence assay with confocal laser scanning microscopy indicated that the ORF136 protein was distributed in the cytoplasm of CCB cells infected with CyHV-3 and transfected with a pVAX1-ORF136 plasmid. Furthermore, immunogold electron microscopy confirmed that ORF136 protein localized to the CyHV-3 envelope.

Development of a double‐antibody sandwich enzyme‐linked immunosorbent assay (DAS‐ELISA) for the detection of KHV

“…Forty proteins of KHV were identified using mass spectrometry (Michel et al., 2010), and the other six via comprehensive proteomic approaches were confirmed (Yi et al., 2014). A total of 16 envelope proteins, 2 tegument proteins and 3 capsid proteins in 46 proteins have been identified (Michel et al., 2010; Rosenkranz et al., 2008; Vancsok et al., 2017; Yi et al., 2014; Zheng et al., 2017). The ORF132 protein was predicted by proteomic analysis to be a membrane protein.…”

“…The ORF132 protein was predicted by proteomic analysis to be a membrane protein. The KHV antigenic proteins that have been characterized are ORF68 and ORF136 (Aoki et al., 2007; Zheng et al., 2017). The ORF25 and ORF81 proteins are the structural proteins, and mainly are located in the cytoplasm, both are immunogenic proteins of KHV, and the constructed DNA vaccines that contain ORF25 and ORF81 genomic fragments could well protect fish in immunized group (Zhou, Wang, et al., 2014; Zhou, Xue, et al., 2014).…”