Articles

Recombinant toxin-coregulated pilus A (TcpA) as a candidate subunit cholera vaccine

Abstract

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.

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IssueVol 6 No 2 (2014) QRcode
SectionArticles
Keywords
Escherichia coli Recombinant Toxin co-regulated pilus Vaccine Vibrio cholerae

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How to Cite
1.
Kiaie S, Abtahi H, Mosayebi G, Alikhani MY, Pakzad I. Recombinant toxin-coregulated pilus A (TcpA) as a candidate subunit cholera vaccine. Iran J Microbiol. 1;6(2):68-73.