Expression of Recombinant Envelope Proteins VP19 and VP28 of WSSV and Preparation of Neutralizing Antibody
Abstract: Two pairs of primers were designed according to the sequences of envelope genes, vp19 and vp28 of White spot syndrome virus (WSSV) in the GenBank. The two DNA fragments about 370bp and 630bp amplified by PCR were linked to the EcoR I restriction endonuclease site and cloned into E.coli expression vector pET- 22b(+) in the proper Open Reading Frame(ORF). With IPTG induction at 35℃, the molecular weight of the engineered protein was about 41kDa, which was identified by SDS-PAGE analysis. Antiserum of the fusion envelope protein VP (19+28) purified by Ni2+- column chromatography was prepared by immunizing rabbit. The VP (19+28) antiserum was used to neutralize the WSSV infection on crayfish by intramuscular injection. The result showed that the antiserum was effective in the neutralization of WSSV on the crayfish which were reared with artificial food at 15-22℃.