Infectious spleen and kidney necrosis virus (ISKNV) is the causative agent of a disease causing high mortality and economic losses in Chinese perch, Siniperca chuatsi in China. Little information about the antigenicity of ISKNV proteins is available. In this study the ORF093 gene of ISKNV was cloned into the prokaryotic expression vector pET32a(+) and eukaryotic expression vector pcDNA3.1(+), and designated as pET-093 and pcDNA-093, respectively. A recombinant 51-kDa protein was detected in Escherichia coli BL21 (harboring pET-093) after IPTG inducement. Polyclonal antibodies were raised in rabbits against the purified ORF093 protein and the reaction of the antibodies was confirmed by western blotting using the purified recombinant protein. Expression of the pcDNA-093 in muscle tissue of vaccinated fish was confirmed by western blotting. The protection efficacy of ORF093 was investigated and results showed that cumulative mortality of Chinese perch was significant differences between immune groups and control(P < 0.05) after ISKNV challenge, and the RPS value of 093 recombinant protein, pcDNA093 and pcDNA093 + MCP was 47%, 50% and 57%. The results suggested that ORF093 is an effective vaccine candidate for ISKNV and it can be used in the control of ISKNV disease in Chinese perch.