Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 6 2 2014 04 15 68 73 EN Somayeh Kiaie Department of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, Iran. Hamid Abtahi Department of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, Iran AND Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran. Ghasem Mosayebi Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran. Mohammad Yosef Alikhani Department of Microbiology, School of Medicine, Hamedan University of Medical Sciences, Hamedan, Iran. Iraj Pakzad Clinical Microbiology Research Center, Ilam University of Medical Sciences, Ilam, Iran AND Department of Microbiology, School of Medicine, Ilam University of Medical Sciences, Ilam, Iran. 2015 10 16 Background   and   Objectives:   The   toxin   co-regulated   pilus   A   (TcpA)   has   been   described   as   a   criti- cal  pathogenicity  factor  of  Vibrio  cholerae.  TcpA  is  a  candidate  for  making  subunit  vaccine  against  chol- era.  The  aim  of  this  study  was  to  produce  a  candidate  vaccine  by  expressing  recombinant  TcpA  in  E.  coli. Materials and Methods: In this study, the toxin co-regulated pilus A gene from EL-Tor, V. cholerae subspecies, was am- plified 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 in- tegrity of product was confirmed by Western blot analysis using a standard rabbit anti V. cholerae antibody. Sera re- activity  of  infected  individual  was  further  analyzed  against  the  recombinant  TcpA  protein.  The  obtained  data  in- dicated  that  recombinant  TcpA  protein  from  V.  cholerae  was  recognized  by  patient  serum  and  animal  sera. Conclusion: These results show that the recombinant TcpA is antigenic and could be used in a carrier host as an oral vaccine against cholera. https://ijm.tums.ac.ir/index.php/ijm/article/view/453 https://ijm.tums.ac.ir/index.php/ijm/article/download/453/230