CLONING OFFIMH AND FLIC AND EXPRESSION OF THE FUSION PROTEIN FIMH/FLIC FROM UROPATHOGENICESCHERICHIA COLI (UPEC) ISOLATED IN IRAN

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ASADI KARAM M.R.; OLOOMI M.; HABIBI M.; BOUZARI SAIED

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Journal Paper

Paper Information

Journal: Iranian Journal of Microbiology Year:2012 | Volume:4 | Issue:2 Page(s): 55-62

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Information Journal Paper

Title

CLONING OFFIMH AND FLIC AND EXPRESSION OF THE FUSION PROTEIN FIMH/FLIC FROM UROPATHOGENICESCHERICHIA COLI (UPEC) ISOLATED IN IRAN

Author(s)

ASADI KARAM M.R. | OLOOMI M. | HABIBI M. | BOUZARI SAIED | Issue Writer Certificate

Keywords

URINARY TRACT INFECTION

UROPATHOGENIC ESCHERICHIA COLI

FIMH

FLIC

FUSION PROTEIN EXPRESSION

Abstract

Background and Objectives: URINARY TRACT INFECTION (UTI) is one of the most common infections in the world. The majority of UTIs are caused by UropathogenicEscherichia coli (UPEC) strains. FIMH and FLIC are the most important virulence factors of UPEC. To date, any ideal vaccine against UTI has not been approved for human use and we need to test new targets to develop an ideal vaccine against UTI. In this study, we constructed fusion FIMH/fliC of UPEC as a novel vaccine candidate against UTI.Material and Methods: PCR amplification of FIMH and FLIC genes of the UPEC isolates was performed by specific primers designed for this purpose. Construction of FIMH/fliC hybrid gene was performed by overlap PCR. The FIMH, FLIC and FIMH/ FLIC were cloned in pET28a vector. The confirmation of expression of the proteins was done by SDS-PAGE and Western blot.Results: The FLIC and FIMH genes were amplified in all of the UPEC isolates tested. The FIMH showed significant homology with the sequences in GenBank. We generated a fusion consisting of the FIMH linked to the N-terminal end of FLIC. Sequencing of the fusion FIMH/fliC showed that fusion was constructed correctly. SDS-PAGE and western blot confirmed the expression of the proteins in optimized condition.Conclusion: URINARY TRACT INFECTION is a huge burden on healthcare system in many countries. UPEC is isolated in around 80% of UTI cases. Antibiotic therapy resulted in the emergence of antibiotic resistance in UPEC strains. This is the major cause for an increasing requirement for a vaccine to prevent UTI. This work describes for the first time the construction of a novel fusion protein from Iranian UPEC isolates. Further immunological studies are required for evaluation of this protein as a novel and safe vaccine candidate against UTI caused by UPEC.

Cites

PREVALENCE OF URINARY TRACT INFECTION AMONG PREGNANT WOMEN AND ITS COMPLICATIONS IN THEIR NEWBORNS DURING THE BIRTH IN THE HOSPITALS OF DEZFUL CITY, IRAN, 2012-2013

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APA: Copy

ASADI KARAM, M.R., OLOOMI, M., HABIBI, M., & BOUZARI, SAIED. (2012). CLONING OFFIMH AND FLIC AND EXPRESSION OF THE FUSION PROTEIN FIMH/FLIC FROM UROPATHOGENICESCHERICHIA COLI (UPEC) ISOLATED IN IRAN. IRANIAN JOURNAL OF MICROBIOLOGY, 4(2), 55-62. SID. https://sid.ir/paper/315312/en

Vancouver: Copy

ASADI KARAM M.R., OLOOMI M., HABIBI M., BOUZARI SAIED. CLONING OFFIMH AND FLIC AND EXPRESSION OF THE FUSION PROTEIN FIMH/FLIC FROM UROPATHOGENICESCHERICHIA COLI (UPEC) ISOLATED IN IRAN. IRANIAN JOURNAL OF MICROBIOLOGY[Internet]. 2012;4(2):55-62. Available from: https://sid.ir/paper/315312/en

IEEE: Copy

M.R. ASADI KARAM, M. OLOOMI, M. HABIBI, and SAIED BOUZARI, “CLONING OFFIMH AND FLIC AND EXPRESSION OF THE FUSION PROTEIN FIMH/FLIC FROM UROPATHOGENICESCHERICHIA COLI (UPEC) ISOLATED IN IRAN,” IRANIAN JOURNAL OF MICROBIOLOGY, vol. 4, no. 2, pp. 55–62, 2012, [Online]. Available: https://sid.ir/paper/315312/en

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