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Infection and Immunity, April 2003, p. 2266-2271, Vol. 71, No. 4
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.4.2266-2271.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Expression of Helicobacter pylori Vacuolating Toxin in Escherichia coli

Mark S. McClain1 and Timothy L. Cover1,2,3*

Departments of Medicine,1 Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232,2 Veterans Affairs Medical Center, Nashville, Tennessee 372123

Received 24 October 2002/ Returned for modification 13 December 2002/ Accepted 30 December 2002

VacA is a secreted toxin that plays a role in Helicobacter pylori colonization of the stomach and that contributes to the pathogenesis of peptic ulcer disease. Studies of VacA structure and function have been hindered by the lack of an efficient system for expression and genetic manipulation of this toxin. In this study, we developed methodology for expression of a functionally active VacA toxin in Escherichia coli. We then used a high-throughput screen to analyze a library of mutant toxins with pentapeptide insertions and identified six mutants that lacked the capacity to induce vacuolation of HeLa cells. The capacity to analyze VacA in this heterologous-expression system should greatly facilitate efforts to elucidate the structure and function of this toxin.

* Corresponding author. Mailing address: Division of Infectious Diseases, A3310 MCN, Vanderbilt University School of Medicine, Nashville, TN 37232. Phone: (615) 322-2035. Fax: (615) 343-6160. E-mail: timothy.l.cover{at}vanderbilt.edu.

Editor: J. T. Barbieri

Infection and Immunity, April 2003, p. 2266-2271, Vol. 71, No. 4
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.4.2266-2271.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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