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Cloning, expression, purification, and characterization of recombinant flagellin isolated from Pseudomonas aeruginosa

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 infec... Full description

Journal Title: Biotechnology Letters 2009, Vol.31(9), pp.1353-1360
Main Author: Goudarzi, Gholamreza
Other Authors: Sattari, Morteza , Roudkenar, Mehryar , Montajabi-Niyat, Mehran , Zavaran-Hosseini, Ahmad , Mosavi-Hosseini, Kamran
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: ISSN: 0141-5492 ; E-ISSN: 1573-6776 ; DOI: 10.1007/s10529-009-0026-1
Link: http://dx.doi.org/10.1007/s10529-009-0026-1
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recordid: springer_jour10.1007/s10529-009-0026-1
title: Cloning, expression, purification, and characterization of recombinant flagellin isolated from Pseudomonas aeruginosa
format: Article
creator:
  • Goudarzi, Gholamreza
  • Sattari, Morteza
  • Roudkenar, Mehryar
  • Montajabi-Niyat, Mehran
  • Zavaran-Hosseini, Ahmad
  • Mosavi-Hosseini, Kamran
subjects:
  • fliC
  • Inclusion bodies
  • Pseudomonas aeruginosa
  • Recombinant flagellin
ispartof: Biotechnology Letters, 2009, Vol.31(9), pp.1353-1360
description: 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 Ni 2+ -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.
language: eng
source:
identifier: ISSN: 0141-5492 ; E-ISSN: 1573-6776 ; DOI: 10.1007/s10529-009-0026-1
fulltext: fulltext
issn:
  • 1573-6776
  • 15736776
  • 0141-5492
  • 01415492
url: Link


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titleCloning, expression, purification, and characterization of recombinant flagellin isolated from Pseudomonas aeruginosa
creatorGoudarzi, Gholamreza ; Sattari, Morteza ; Roudkenar, Mehryar ; Montajabi-Niyat, Mehran ; Zavaran-Hosseini, Ahmad ; Mosavi-Hosseini, Kamran
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descriptionPseudomonas 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 Ni 2+ -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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titleCloning, expression, purification, and characterization of recombinant flagellin isolated from Pseudomonas aeruginosa
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titleCloning, expression, purification, and characterization of recombinant flagellin isolated from Pseudomonas aeruginosa
authorGoudarzi, Gholamreza ; Sattari, Morteza ; Roudkenar, Mehryar ; Montajabi-Niyat, Mehran ; Zavaran-Hosseini, Ahmad ; Mosavi-Hosseini, Kamran
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abstractPseudomonas 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 Ni 2+ -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.
copDordrecht
pubSpringer Netherlands
doi10.1007/s10529-009-0026-1
pages1353-1360
date2009-09