›› 2010, Vol. 37 ›› Issue (2): 83-86.

• 生物技术 • Previous Articles     Next Articles

Prokaryotic Expression of Infectious Bovine Rhinotracheitis Virus Recombinant Truncated Glycoprotein B and Development of Indirect ELISA for Detection of it’s Antibody

LI Wei1，3, MENG Ri-zeng2, SHI Jian-ping2, HE Jiang-shuai1, ZHAO Xiao1， LU Qiang1   

  1. （1.Institute of Zoonoses, Jilin University, Changchun 130062, China;2.Jilin Entry-Exit Inspection and Quarant Ine Bureau, Changchun 130062, China；3.Heilongjiang Vocational College of Biology Science and Technology, Harbin 150025, China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-02-20 Published:2010-02-20

Abstract: Abstract: The hydrophilicity, antigenic index and surface probability of glycoprotein B (gB) of infectious bovine rhinotracheitis virus (IBRV) were analyzed by biosoftware DNAStar-Protean. Amplified the sequence which codes the amino acid residues from 385th to 550th in major antigen determinant region of gB glycoprotein. Then the purified products were cloned into pMD18-T vector and sequenced. The truncated gB gene was inserted into pET-28b vector which was transformed to competent Escherichia coli DE3 for expressing. The SDS-PAGE showed that the fusion protein of 21.35 ku as expected. Western blotting and indirect ELISA analysis that the recombinant protein had a specific positive reaction with standard positive blood serum. The concentration of purified protein was 2.0 mg/mL and the purity was 95.27%. The preliminary indirect ELISA method was developed by using the purified recombinant protein of gB as antigen. The results above suggested that the assay was confirmed to be specific, sensitive and quickly, which can be used as antigen for diagnosis.

Key words: Key words: infectious bovine rhinotracheitis virus; glycoprotein B; prokaryotic expression; indirect ELISA