African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12296

Full Length Research Paper

Expression of infectious bovine rhinotracheitis virus glycoprotein D in bacterial cell

Hassan Momtaz1* and Mohammadreza Nazem2
1Department of Microbiology, Faculty of Veterinary Medicine, Islamic Azad University, Shahrekord Branch- Shahrekord- Iran. 2Faculty of Veterinary Medicine, Islamic Azad University, Shahrekord Branch- Shahrekord- Iran.
Email: [email protected]

  •  Accepted: 23 March 2010
  •  Published: 14 June 2010

Abstract

    

Bovine Herpesvirus 1 (BHV-1) belongs to the genus of Varicellovirus and the family of Herpesviridae which contains three main gB, gC and gD genes. In order to cloning of the coding region of gD gene of IBR virus , PCR product of the open reading frame of the gene from IBR virus isolated in Iran was amplified by PCR. A 1047bp PCR product of the gD gene with EcoRI, HindIII restriction sites were subcloned of pTZ57R/T and digested by the mentioned endonucleases. Digested insert cloned in to pET-32a and transfered in E.coli cells. For the expression of gD protein, the pET-32a recombinant vector was transformed and then induced in BL21 (DE3) strain of E.coli competent cells using IPTG. The presence of gD expressed protein was shown in immunoblotting and SDS-PAGE system. With respect to the remarkable frequency of infection to IBR in Iran and the necessity of controlling it through vaccination with recombinant vaccines of thymidine kinase, manufacturing and applying the recombinant gD protein are vital goals in recognition and distinction between infection and responses caused by vaccine.

 

Key words: IBR virus, gD protein, pET-32a vector, protein expression, SDS-PAGE, immunoblotting.

    

 

 

 

 

Abbreviation

BHV-1, Bovine herpesvirus 1; IBR, infectious bovine rhinotracheitis; gD, glycoprotein D; IPTG, isopropyl β-D-1-thiogalactopyranoside;SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis; DMSO,dimethylsulfoxide; PVDF, polyvinylidine difluoride; DAB, diamino benzidine; BSA,bovine serum albumin; ELISA, enzyme-linked immunosorbent assay; TBST, tris buffer saline Tween 20.