Foot-and-mouth disease (FMD) is an acute, highly contagious disease that infects cloven-hoofed animals. There are seven serotypes of FMD virus (FMDV), namely O, A, C, SAT1, SAT2, SAT3 and Asia1.Serotypes A, O and SAT2, are responsible for the epidemics in our country and we are selected SAT2 as model in this work. In this study we are discuss the influence of time and concentration of isopropyl-β-D-thiogalactoside (IPTG) for induction of the tac-promoter as a synthetically produced DNA promoter, in small-scale cultivations is well established for over gene expression using Synthetic gene of recombinant 1B Capsid Protein of FMDV. For this purpose, the 1B-coding sequence was modified and optimized to match the codon usage preference of E. coli. The modified 1B–coding sequence was synthesize, amplification and cloning to construct of1B-PGEX4T1 bacterial expression vector and transformation in to E. coli BL21 (DE3) strain as expression host induction of the gene led to expression of the target protein. It is worthy that the best condition for 1B protein expression was 0.1mM IPTG at 16°C for overnight incubation. The expressed protein was analyzed by SDS-PAGE and the presence of a band with a molecular mass of approximately 53 KDa (the expected size for the GST tagged protein 26 KDa and 1B recombinant protein 22 KDa) and the yield of recombinant 1B protein was 1mg/100ml of culture medium. |
