Expression and purification of recombinant immunogenic protein containing Shigella dysentery Virulence Factors

Shigella dysentery is a biological agent that causes bloody diarrhea. Making an effective immunogen against bacteria is essential by emergence of antibiotic resistance . Binding, invasive and toxin factors are one of the most important vaccine candidates against Shigella dysentery. Accordingly, the aim of this study was to express recombinant Immunogenic Chimeric with Shigella Dysenteric Virulence Factors.

Chimer gene coder optimization was performed with OPTIMIZER software. The gene constructs were cloned in pET32a vector. The recombinant plasmid was transferred to E. coli BL21 DE3 cells and expression of the recombinant protein was induced using IPTG. The recombinant protein was purified by chromatography and was evaluated with western blotting.

The codon compatibility index (CAI) for the natural gene was 0.67, while the optimized gene had an index of 0.9. The enzymatic analysis confirmed the accuracy of the chimer gene cloning in the vector. The expression of recombinant protein in E. coli caused the production of a recombinant protein of 80 kDa. Western blotting showed the reaction of recombinant protein with anti-histidine antibody. The amount of purified protein of the culture medium was 2.5 mg/ml.

The expression of the recombinant chimeric protein with 80 kDa in E.coli host and its purification was successfully carried out. The recombinant chimeric protein can be injected or loaded onto nanoparticles to evaluate oral and injectable immunizations.