Veterinary Research Forum
Vet Res Forum

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

Document Type : Original Article

Authors

Fereshteh Yazdani 1
Mehran Bakhshesh 1
Majid Esmaelizad 2
Zohre Azita Sadigh 3

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

BEFV
Escherichia coli
ELISA
G1- Epitope

Subjects

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Veterinary Research Forum
Volume 8, Issue 3
September 2017
Pages 209-213

  • Receive Date 15 September 2017

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