کلون و بیان ژن fliC به‌عنوان کاندیدای واکسن علیه بیماری تیفوئید

Background & Aims: Salmonella enterica is a zoonotic pathogen causing typhoid fever in humans and animals. It is an important cause of food borne infections in humans throughout the world. A recent study estimated approximately 22 million cases of typhoid each year with at least 200,000 deaths. FliC encoding flagellin plays a role in pathogenesis and is an important antigen in vaccination. In this study, recombinant protein fliC as a candidate vaccine against salmonellosis was produced and purified. Materials & Methods: The protein sequence of fliC was obtained from NCBI database. For high level expression of protein, the gene was synthesized with codon bias of E. coli.; synthetic gene in pUC57 was sub coloned into pET32 expression vector and then transferred into E. coli BL21DE3. The recombinant protein was express by IPTG induction. For high expression, three parameters including IPTG concentration, time and temperature of induction were optimized. The recombinant protein was purified by ion exchange chromatography with His-Tag. The purified protein was confirmed by western blotting. Results: Recombinant vector was approved by PCR and restriction analysis. SDS-PAGE showed a 51 kDa protein with proper purity and western blotting with his-tag antibody confirmed the intend protein. Conclusion: The use of recombinant vaccines is highly regarded, since one of the major virulence factors of Salmonella gene was cloned and the protein was produced and purified in this study. This protein can be used in candidate vaccine against typhoid. SOURCE: URMIA MED J 2016: 27(8): 691 ISSN: 1027-3727