Abstract: The ORF8 sequence of Anguillid herpesvirus (AngHV) was amplified from the genome of AngHV-FJ strain by PCR with primers based on the sequence of AngHV reference strain (NC_013668) in GenBank, and then the target fragment was cloned into pMD19-T vector and verified by restriction enzyme digestion and sequencing. Bioinformatics analysis indicated that ORF8 protein might be anchor on the viral envelop with good stability, transmembrane structure, and numbers of antigen epitopes. The analysis of transcription profiles revealed that ORF8 gene was the early gene of AngHV and probably had an active role in the early stage of viral infection. ORF8 gene was then cloned into the expression vector pET-32a and transformed into Escherichiacoli BL21 (DE3). The encoded protein was highly expressed under the induction of IPTG and followed by SDS-PAGE and Western blot verification. The expressed high-purity ORF8 protein with relative molecular mass of 44 000 was obtained by following gel extraction purification. The findings provide fundamental data on evaluation of the immunity effect of the expressed protein and preparation of specific ORF8 antibody, as well as on development of immunodiagnostic test reagent and vaccine preparation of the virus.
2021, Vol. 36 
