Abstract:

Koi herpesvirus (KHV) is an emerging pathogen causing mass mortality in koi. the conserved ORF1 gene of KHV was selected to amplification by PCR. The full length of this gene is 774 bp encoding an predicted envelope protein VP1 which consists of 258 amino acids. The amplified gene was subcloned into pET-32a vector to construct recombinant expression plasmids named K1-PET, which then was transformed into E.coli BL21 (DE3) and expressed by IPTG inducement. SDS-PAGE and Western blot results show that K1-PET can be expressed highly. The molecular weight of fusion VP1 protein is 37.3 KD and the protein is mainly expressed as inclusion body. The rabbit serum obtained after four immunizations with the fusion VP1 protein, was then used for ELISA analyses. The result showed that the potency of anti-serum was 1:27000, and the VP1 protein had high immunogenicity. Therefore, the VP1 protein and anti-serum could be used in research of KHV detection method, vaccine produce and protein function study.