Abstract: Anguillae herpesvirus(AngHV)is one of the major viral pathogen of freshwater eels. In this study, the open reading frame of AngHV-FJ ORF95 gene was synthesized according to the gene sequence of AngHV-1 provided by GenBank:(NC-013668),and then cloned into the pMD19-T vector. After restriction digestion and sequence verification,the ORF95 gene was inserted into the prokaryotic expression vector pET-32a to construct the recombinant expression plasmid 32a-ORF95. The plasmid was then transformed into E. coli BL21 (DE3) strain to induce expression of ORF95 protein. The soluble ORF95 protein can be highly expressed under the induction of IPTG (0.1 mmol·L-1) at 23℃ for 14h,and then purified by Ni-NTA His.Binding affinity purification. The protein can be used further to understand the characteristic of ORF95 protein and the AngHV prevention.