Cloning and expression of ipaC gene from Shigella dysenteriae

Shigellosis is caused by Shigella species. Considering the high frequency of morbidity and mortality reports and antibiotics resistance, designing and producing a vaccine against this disease is one of the goals of World Health Organization. Invasion Plasmid Antigen especially IpaC and IpaB are the major Shigella virulence agents and are also vaccine candidates. The aim of this study was to express ipaC gene which is of the virulence factors in order to investigate its immunogenicity. In this study, ipa C gene was obtained from NCBI gene bank and primers were designed. After genome extraction from Shigella dysenteriae, it was used as template for PCR amplification. The amplified ipa C gene by PCR was cloned into pTZ57R and sub-cloned into the expression vector pET-28a(+). E. coli BL21(DE3)plysS was transformed by recombinant vector pET-28a(+)/ ipa C and expression of the recombinant ipa C was investigated by IPTG induction and SDS-PAGE electrophoresis. Cloning was confirmed by PCR, enzyme digestion and sequencing. In addition, the recombinant protein was expressed by IPTG induction. Protein expression was confirmed by Ni–NTA column, Western-Blotting analysis and SDS-PAGE electrophoresis. Cloning, sub-cloning and expression of ipaC gene are confirmed in the present study. Therefore, this recombinant protein can be used for production of a recombinant vaccine against Shigella dysenteriae in future studies, after immunogenicity assay.