Expression of G1- epitope of bovine ephemeral fever virus in E. coli : A novel candidate to develop ELISA kit

Document Type : Original Article

Authors

1 Department of Animal Virology, Research and Diagnosis, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

2 Department of Biotechnology, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

3 Department of Human Vaccine Production, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Organization Extension (AREEO), Karaj, Iran

Abstract

Bovine ephemeral fever is an acute and arthropod-borne viral disease of cattle and water buffalo which occurs seasonally in most of the world tropical and subtropical regions. The epizootic feature of the disease has been reported in Iran with serious economic consequences. The surface glycoprotein G of bovine ephemeral fever virus (BEFV) is composed of 4 antigenic sites (G1-G4) and plays the main role for eliciting neutralizing antibodies and protective immunity. The G1 – epitope is a linear antigenic site and conserved among BEFV strains. In order to develop an ELISA test based on G1-epitope as coating antigen, this study was carried out to express the recombinant G1-epitope of BEFV in prokaryotic system. Using PCR and specific primers, a length of 88 amino acid of the G glycoprotein of BEFV including G1- epitope was amplified and cloned into the expression vector pGEX-4T-1, with the GST moiety. The recombinant plasmid (pGEX-4T-1-G1) was then transformed into Escherichia coli BL21 and expression of fusion protein was induced by 0.10 mM IPTG. The maximum expression of the fusion protein was obtained at 16 hr post induction as verified by SDS-PAGE electrophoresis, and it was also confirmed that this protein bearing G1- epitope is sufficiently biologically active to bind to anti-BEFV serum in western blot experiment.

Keywords

Main Subjects


  1. Walker PJ. Bovine ephemeral fever in Australia and the world. Curr Top Microbiol Immunol 2005; 292: 57-80.
  2. Bakhshesh M, Abdollahi D. Bovine ephemeral fever in Iran: Diagnosis, isolation and molecular characterization. J Arthropod Borne Dis 2015; 9 (2):195-203.
  3. Dietzgen RG. Calisher CH. Kurath G et al. Rhabdoviridae In: King AMQ, Adams MJ, Carstens EB, et al (Eds). Virus taxonomy. London, UK: Elsevier 2012; 686-713.
  4. Walker PJ, Byrne KA, Cybinski DH, et al. Proteins of bovine ephemeral fever virus. J Gen Virol 1991;72 (1): 67-74.
  5. Cybinski DH, Walker PJ, Byrne KA, et al. Mapping of antigenic sites on the bovine ephemeral fever virus glycoprotein using monoclonal antibodies. J Gen Virol 1990;71 (9): 2065-2072.
  6. Uren MF, Walker PJ, Zakrzewski H, et al. Effective vaccination of cattle using the virion G protein of bovine ephemeral fever virus as an antigen. Vaccine 1994;12 (9):845-852.
  7. Kongsuwan K, Cybinski DH, Cooper J, et al. Location of neutralizing epitopes on the G protein of bovine ephemeral fever rhabdovirus. J Gen Virol 1998; 79 (11):2573-2581.
  8. Zakrzewski H, Cybinski DH, Walker PJ. A blocking ELISA for the detection of specific antibodies to bovine ephemeral fever virus. J Immunol Methods 1992;151 (1): 289-297.
  9. Zheng FY, Lin GZ, Qiu CQ, et al. Expression and antigenic characterization of the epitope-G1 of the bovine ephemeral fever virus glycoprotein in Pichia pastoris. Virol Sin 2007; 22(5):347-352.
  10. Zheng FY, Lin GZ, Qiu CQ. Expression, purification and antigenic characterization of the Epitope-G1 gene of bovine ephemeral fever virus in Escherichia coli [Chinese]. Acta Microbiol Sin 2007; 47: 498-502.
  11. Wang FI, Hsu AM, Huang KJ. Bovine ephemeral fever in Taiwan. J Vet Diagn Invest 2001;13 (6): 462-467.
  12. Aziz-Boaron O, Klausner Z, Hasoksuz M, et al. Circulation of bovine ephemeral fever in the Middle East—strong evidence for transmission by winds and animal transport. Vet Microbiol 2012; 158 (3): 300-307.
  13. Aziz‐Boaron O, Brettschneider S, King R, et al. Seroprevalence of bovine ephemeral fever virus in domesticated and wildlife species during epidemic and inter‐epidemic periods (2000–2009) in Israel. Transbound Emer Dis 2015; 62 (2):183-187.
  14. Finlaison DS, Read AJ, Kirkland PD. An epizootic of bovine ephemeral fever in New South Wales in 2008 associated with long‐distance dispersal of vectors. Aust Vet J 2010;88 (8):301-306.
  15. Yang DK, Kim SY, Kim HH, et al. The follow up study after massive outbreak of Akabane and bovine ephemeral fever viruses in Korea. Korean J Vet Serv 2013; 36 (3):151-155.
  16. Niwa T, Shirafuji H, Ikemiyagi K, et al. Occurrence of bovine ephemeral fever in Okinawa Prefecture, Japan, in 2012 and development of a reverse-transcription polymerase chain reaction assay to detect bovine ephemeral fever virus gene. J Vet Med Sci 2015; 77(4):455-460.
  17. Ting LJ, Lee MS, Lee SH, et al. Relationships of bovine ephemeral fever epizootics to population immunity and virus variation. Vet Microbiol 2014; 173 (3): 241-248.
  18. Cybinski DH. Homologous and heterologous antibody reactions in sera from cattle naturally infected with bovine ephemeral fever group viruses. Vet Microbiol 1987; 13(1): 1-9.
  19. Zheng FY, Lin GZ, Qiu CQ, et al. Serological detection of bovine ephemeral fever virus using an indirect ELISA based on antigenic site G 1 expressed in Pichia pastoris. Vet J 2010; 185 (2): 211-215.
  20. Zheng FY, Lin GZ, Qiu CQ, et al. Development and application of G 1-ELISA for detection of antibodies against bovine ephemeral fever virus. Res Vet Sci 2009; 87 (2): 211- 212.