Cloning and Expression of FimH Lectin Domain as a Candidate Immunogen against Urinary Infection

Abstract

Background and Objectives: Urinary tract infection (UTI) is one of the most common infections in human, and Uropathogenic Escherichia coli (UPEC) strains are the most common cause of UTI. These strains colonize in bladder and cause cystitis, which could cause pyelonephritis by movement to the kidneys. Since FimH protein has a very important role in colonization of UPEC strains and causing infection, this study was conducted with the purpose of designing FimH protein as a candidate immunogen against urinary infection. Methods: In the present research, fimH adhesin was selected as a candidate immunogen. Genomic DNA was extracted from E. coli 35218 and fim H gene was amplified by PCR. The PCR product was cloned into pBluescript SK vector and confirmed by sequencing. Then, fim H lectin domain was amplified and inserted into pET23a expression vector. Results: The obtained pET/fimH clone was verified by sequencing and transferred into E. coli origami (DE3) strain. Expression of recombinant FimH protein in E. coli was verified by SDS-PAGE and western blot methods. Conclusion: According to the results of this study, the produced protein could be investigated for evaluation of immunogenicity in animal models.

Keywords

• urinary tract infection
• uropathogenic escherichia coli
• vaccines
• synthetic
• genetic vaccine
• fimh protein
• e coli