Prokaryotic Expression Of Non-Virion Protein Of Hirame Rhabdovirus And Preliminary Analysis Of Its Antigeneicity

Hirame rhabdovirus( HRV) is a Novirhabdovirus resulting in lethal disease in wild and farmed Japanese ounder(Paralichthys olivaceus) with a severe economic losses in aquaculture industry. During the invasion of the virus in the host cell, and the perceived invasion of host cells by the virus pattern recognition receptors monitor pathogen associated molecular patterns, and passed through a cascade of signaling pathways, coordination viral infection early host response, and then activate the host acquired immunity reaction. However, by the interaction of RNA viruses and hosts, RNA viruses evolved to escape immune surveillance of the host RLRs different strategies to interfere with the host RLRs antiviral signaling, blocking signal transduction RLRs, leading host disease. In most cases, the virus tended to use Non-virion perotein to escape immune surveillance of the host RLRs. But the molecular mechanisms of HRV pathogenesis and host immune surveillance escape have not been clearly explained.In this study, according to the full-length gene sequence of HRV Chinese strain in Gen Bank specific primers were designed using RT-PCR method amplified fragment from the viral suspension of HRV. The homology of resulting fragments sequence reached 99.77 percent, and preparing for the Infectious clone.By RT-PCR method, the fragment of NV gene was amplified from HRV Chinese isolate. The NV gene was cloned into the p Cold vectorⅠ,then the recombinant expression plasmid was transformed into E.coli BL21(T1®) and induced by IPTG for the expression of non-virion protein. Purified with Ni-NTA affinity column to acquire NV protein. Western-blot analysis indicated that the anti-serum could recognize the NV protein of HRV. The successful experiment of NV protein was the experimental basis of HRV escape host immune surveillance mechanisms.