title: Recombinant toxin-coregulated pilus A (TcpA) as a candidate subunit cholera vaccine.
abstract: OBJECTIVE
  The toxin co-regulated pilus A (TcpA) has been described as a critical pathogenicity factor of Vibrio cholerae. TcpA is a candidate for making subunit vaccine against cholera. The aim of this study was to produce a candidate vaccine by expressing recombinant TcpA in E. coli.
  
  
  METHODS
  In this study, the toxin co-regulated pilus A gene from EL-Tor, V. cholerae subspecies, was amplified by PCR and sub-cloned into prokaryotic expression vector pGEX4T1. E. coli BL21 (DE3) was transformed with pGEX4T1- TcpA and gene expression was induced by IPTG and purified by GST resin. The integrity of the product was confirmed by Western blot analysis using a standard rabbit anti-V. cholerae antibody. Sera reactivity of infected individuals was further analyzed against the recombinant TcpA protein.
  
  
  RESULTS
  The concentration of purified recombinant protein was calculated to be 8 mg/L of initial culture. The integrity of product was confirmed by Western blot analysis using a standard rabbit anti V. cholerae antibody. Sera reactivity of infected individual was further analyzed against the recombinant TcpA protein. The obtained data indicated that recombinant TcpA protein from V. cholerae was recognized by patient serum and animal sera.
  
  
  CONCLUSIONS
  These results show that the recombinant TcpA is antigenic and could be used in a carrier host as an oral vaccine against cholera.
authors.affiliation: Department of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, Iran.
authors.forename: Somayeh
authors.initials: S
authors.lastname: Kiaie
authors.affiliation: Department of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, Iran ; Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran.
authors.forename: Hamid
authors.initials: H
authors.lastname: Abtahi
authors.affiliation: Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran.
authors.forename: Ghasem
authors.initials: G
authors.lastname: Mosayebi
authors.affiliation: Department of Microbiology, School of Medicine, Hamedan University of Medical Sciences, Hamedan, Iran.
authors.forename: MohammadYosef
authors.initials: M
authors.lastname: Alikhani
authors.affiliation: Clinical Microbiology Research Center, Ilam University of Medical Sciences, Ilam, Iran ; Department of Microbiology, School of Medicine, Ilam University of Medical Sciences, Ilam, Iran.
authors.forename: Iraj
authors.initials: I
authors.lastname: Pakzad
keywords: Escherichia coli; Recombinant; Toxin co-regulated pilus; Vaccine; Vibrio cholerae