Cloning and Expression of N-teminal Domain of Pseudomonas aeruginosa Flagellin and Evaluation of Antibodies Raised against it on Motility Inhibition of Pseudomonas aeruginosa

سال انتشار: 1391
نوع سند: مقاله ژورنالی
زبان: انگلیسی
مشاهده: 250

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شناسه ملی سند علمی:

JR_ZUMS-20-80_001

تاریخ نمایه سازی: 11 اردیبهشت 1400

چکیده مقاله:

Background and Objective: Pseudomonas aeruginosa is an opportunistic pathogen that causes severe and lethal infections in immunocompromised individuals. This bacterium possesses a single polar flagellum. Flagellum and its subunit Flagellin play important roles in the pathogenesis of P. aeruginosa. Flagellin induces immune responses by interaction of its N-terminal domain with TLR-۵. Our main aims of this study were cloning and expression of N-terminal domains of flagellin and evaluation of antibodies raised against it on motility inhibition of P. aeruginosa. Material and Methods: The DNA sequence coding for the first ۱۶۱ amino acids of flagellin was PCR amplified and cloned into a pET-۲۸a expression vector. Recombinant protein was over expressed in BL-۲۱(DE۳), and purified by Ni-NTA resin. The immune reactivity of recombinant truncated flagellin was evaluated by Western blotting. The recombinant protein was injected into a rabbit and antibodies raised against it were evaluated for the cell motility inhibition of P. aeruginosa ۸۸۲۱M. Results: The N-terminal domain of Flagellin was successfully overexpressed in Escherichia coli BL-۲۱(DE۳) host strain. Anti-native and anti-N-terminal flagellin antibodies reacted with the recombinant protein. Motility inhibition assay demonstrated that polyclonal antiserum against N-teminal flagellin is able to inhibit the motility of P. aeruginosa ۸۸۲۱M. Conclusion: The N-terminal domain of flagellin may be used for development of a new recombinant vaccine against P. aeruginosa infections.

کلیدواژه ها:

Keywords: Cloning ، Motility inhibition ، N-terminal domains of flagellin ، Pseudomonas aeruginosa

نویسندگان

فریده دکتر زاده

Dept. of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

اشرف محبتی مبارز

Dept. of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

مهریار حبیبی رود کنار

Research Center, Iranian Blood Transfusion Organization, Tehran, Iran

مهدی فروزنده مقدم

Dept. of Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.