Molecular Cloning and Expression of Bovine Viral Diarrhea Virus Nonstructural Protein 3 in Escherichia coli
| Author | Pezhman Mahmoodi | en |
| Author | Masoud Reza Seyfi Abad Shapouri | en |
| Author | Masoud Ghorbanpour | en |
| Author | Mohammad Rahim Haji Hajikolaei | en |
| Author | Mohsen Lotfi | en |
| Author | Maryam Ekhtelat | en |
| Issued Date | 2013-09-01 | en |
| 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. | en |
| DOI | https://doi.org/10.5812/jjm.7044 | en |
| Keyword | Bovine Viral Diarrhea | en |
| Keyword | Nonstructural Protein 3 | en |
| Keyword | Cloning | en |
| Keyword | Expression | en |
| Keyword | <i>Escherichia coli</i> | en |
| Publisher | Brieflands | en |
| Title | Molecular Cloning and Expression of Bovine Viral Diarrhea Virus Nonstructural Protein 3 in Escherichia coli | en |
| Type | Research Article | en |
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