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Biotechnology Letters

, Volume 31, Issue 9, pp 1353–1360 | Cite as

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

  • Gholamreza Goudarzi
  • Morteza Sattari
  • Mehryar Habibi Roudkenar
  • Mehran Montajabi-Niyat
  • Ahmad Zavaran-Hosseini
  • Kamran Mosavi-Hosseini
Original Research Paper

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.

Keywords

fliC Inclusion bodies Pseudomonas aeruginosa Recombinant flagellin 

Notes

Acknowledgments

We thank Professor Ian Alan Holder (Departments of Microbiology and Surgery, University of Cincinnati College of Medicine, Cincinnati, Ohio 45267) for his scientific support and revising of the manuscript.

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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Gholamreza Goudarzi
    • 1
  • Morteza Sattari
    • 1
  • Mehryar Habibi Roudkenar
    • 2
  • Mehran Montajabi-Niyat
    • 1
  • Ahmad Zavaran-Hosseini
    • 3
  • Kamran Mosavi-Hosseini
    • 2
  1. 1.Department of Bacteriology, Medical Sciences SchoolTarbiat Modares UniversityTehranIran
  2. 2.Research Center, Iranian Blood Transfusion OrganizationTehranIran
  3. 3.Department of Immunology, Medical Sciences SchoolTarbiat Modares UniversityTehranIran

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