Cloning,expression, purification,and characterization of recombinant flagellin isolated from <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> |
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| Authors: | Gholamreza Goudarzi Morteza Sattari Mehryar Habibi Roudkenar Mehran Montajabi-Niyat Ahmad Zavaran-Hosseini Kamran Mosavi-Hosseini |
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| Institution: | (1) Department of Bacteriology, Medical Sciences School, Tarbiat Modares University, Tehran, Iran;(2) Research Center, Iranian Blood Transfusion Organization, Tehran, Iran;(3) Department of Immunology, Medical Sciences School, Tarbiat Modares University, Tehran, Iran |
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| Abstract: | Pseudomonas aeruginosa as an opportunistic pathogen causes lethal infections in immunocompromised individuals. This bacterium possesses a polar
flagellum made up of flagellin subunits. Flagella have important roles in motility, chemotaxis, and establishment of P. aeruginosa in acute phase of infections. Isolation, cloning, and expression of flagellin were aimed at in this study. Flagellin gene
(fliC) of P. aeruginosa strain 8821M was isolated by PCR and cloned into a pET expression vector. The recombinant flagellin (46 kDa) was overexpressed
as inclusion bodies (IBs). IBs were solubilized in guanidine hydrochloride (GuHCl) followed by affinity-purification and renatured
using Ni2+-Sepharose resin. Recombinant flagellins reacted with the serum from a rabbit previously immunized with native flagellin.
In addition, polyclonal antiserum raised against the recombinant flagellin was shown to significantly inhibit the cell motility
of P. aeruginosa strain 8821M in vitro. |
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| Keywords: | fliC Inclusion bodies Pseudomonas aeruginosa Recombinant flagellin |
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