Antiserum preparation for infectious hematopoietic necrosis virus (IHNV) glycoprotein and its application in IHNV detection

Abstract:

The 1527bp IHNV glycoprotein gene was cloned by reverse transcription polymerase chain reaction（RT-PCR）from cell culture, and was subcloned into pCWori plasmid. Then the recombinant pCWori-G was transformed into DH5α. The recombinant protein was obtained through fermentation. The 57kDa target protein was expressed successfully by 1 mmol/L IPTG induction for 5 h. SDS-PAGE analysis showed that it was mainly in the form of inclusion body. The soluble glycoprotein was obtained after purification and renaturation processing. Western blot analysis showed that the recombinant protein was identified specifically by the goat anti IHNV serum. The recombinant protein was used to immunize mice to produce glycoprotein antiserum. The prepared antiserum reacted specifically with IHNV by indirect ELISA test. In this study, the IHNV glycoprotein was expressed by prokaryotic expression system and polyclonal antiserum was obtained by immunized mice.