Cloning and Expression of vp1 gene of Foot- and- mouth Disease Virus and fliCgeneof Salmonella in Escherichia coli

Foot& mouth disease (FMD) is a vesicular viral disease which cause severe damage in livestock industry. Today FMD is controlled by traditional inactivated vaccine. Due to short duration of immunity, using molecular adjuvant is recommended to increase efficiency of vaccine. Purpose of this study is synthesis fusion protein from VP1 protein of FMDV (Opanasia2) and flagelline of the Salmonella (fliC) in E. coli. 1D gene that is responsible for producing VP1 protein was produced by PCR (800bp)and cloned in PTZ57 vector. After cutting cloned fragment from the recombinant plasmid by HindIII&XhoI, inserted to expression vector (pET41b) that containsfliC of the Salmonella. The construct was transfected to competent E. coli (BL21DE3) cell. The expression was induced by IPTG (0.5mM) and purified by Ni-NTA Resin. Production of recombinant protein was resolved by SDS-PAGE in 12% acrylamide gel and western blotting. Recombinant fusion protein was evaluated by PCR and enzyme digestion. By molecular analysis, PCR product by length 700 and 1500 bp was observed. Ni-NTA Resin and gradient pH was used for purification. The most amount of protein recovering was observed in pH 4.5. Recombinant protein by molecular weight of 76 KDa determined in SDS-PAGE (12% acrylamide gel) and was confirmed by western blot by AntiHis. In this study, the fused gene of FMDV vp1 and flagelline of Salmonella (fliC/2200 bp) was cloned successfully in E.coliand recombinant protein (76 KDa) was expressed and purified.