molecular cloning and expression of bovine viral diarrhea virus nonstructural protein 3 in escherichia coli
نویسندگان
چکیده
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.
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عنوان ژورنال:
jundishapur journal of microbiologyجلد ۶، شماره ۷، صفحات ۰-۰
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