Author/Authors :
Mahmoodi، Pezhman نويسنده Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, IR Iran , , Seyfi Abad Shapouri، Masoud Reza نويسنده Department of Pathobiology, School of Veterinary Medicine, Shahid Chamran University, P.O. Box 61355-145, Ahvaz, I.R. Iran , , Ghorbanpour، Masoud نويسنده Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, IR Iran , , Haji-Hajikolaei، Mohammad Rahim نويسنده Department of Large Animal Internal Medicine, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran , , Lotfi، Mohsen نويسنده Razi Vaccine and Serum Research Institute, Karaj, IR Iran , , Ekhtelat، Maryam نويسنده Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, IR Iran ,
Abstract :
Background: Bovine viral diarrhea (BVD) is an economically important disease of cattle with a worldwide distribution. Diagnosis of BVD relies on laboratory-based detections of its viral causing agent or virus specific antibodies. The most common laboratory method used for this purpose is the ELISA. Bovine viral diarrhea virus (BVDV) nonstructural protein 3 (NS3) is one of the most highly conserved immunogenic proteins of BVDV, thus, it is a proper candidate antigen to detect antibodies against the virus in the sera from infected animals.
Objectives: The aim of this study was to synthesize a plasmid construct for high-level expression of NS3 with more solubility in Escherichia coli.
Materials and Methods: A segment of BVDV genome encoding the NS3 protein was amplified using RT-PCR and cloned into pMAL-c2X expression vector, under the control of the lac promoter. After sequencing of the amplified gene, the recombinant protein was expressed in E. coli strain BL21 and analyzed by SDS-PAGE and western blotting.
Results: The strong promoter of pMAL-c2X vector allowed a high level expression of NS3 as a maltose binding protein-NS3 (MBP-NS3) fusion protein. Expression of the expected fusion protein was confirmed by electrophoresis on SDS-PAGE and immunoblotting, using a BVDV positive bovine serum.
Conclusions: Based on our results, it appears that this plasmid construct may be suitable for the production of NS3 recombinant antigen to develop BVDV laboratory diagnostic assays.
