The prokaryotic expression and application of epitope-rich region of in infectious hematopoietic necrosis virus

The genomic RNA extracted from infectious hematopoietic necrosis virus Sn1203( IHNV-Sn1203) was used as a template to clone truncated glycoprotein by one-step RT-PCR. The gene which is about 375 bp was cloned into the expression vector p ET-27 b,and the recombinant plasmid p ET-27b-IHNV-short G was highly expressed by E. coli strain Rosetta.The result of SDS-PAGE showed that the protein with expected size 14 k Da was obatained after induced for 4 hours at the IPTG concentration of 0. 25 mmol / L at 37 ℃,and the protein was expressed as the form of inclusion body. The truncated protein was purified by denaturation and renaturation treat using different concentrations of urea,and the protein was obtain without any labels. The purified protein was used to produce antisera. The antisera against glycoprotein prepared in this study could react specifically with both natural glycoprotein of the IHNV-Sn1203 and the recombinant truncated glycoprotein in ELISA test,and the antisera titer against the natural glycoprotein was1∶ 40 000,the recombinant glycoprotein was 1∶ 80000. IFA result showed that the antisera could specifically recognize the glycoprotein on the surface of IHNV-Sn1203 and have no cross-react with VHSV.