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.

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.

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