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Applied and Environmental Microbiology, February 2002, p. 928-932, Vol. 68, No. 2
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.02.928-932.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Alkaline Phosphatase Reporter Transposon for Identification of Genes Encoding Secreted Proteins in Gram-Positive Microorganisms

Carmela M. Gibson and Michael G. Caparon*

Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 63110-1093

Received 2 July 2001/ Accepted 16 October 2001

We describe the construction of TnFuZ, a genetic tool for the discovery and mutagenesis of proteins exported from gram-positive bacteria. This tool combines a transposable element (Tn4001) of broad host range in gram-positive bacteria and an alkaline phosphatase gene (phoZ) derived from a gram-positive bacterium that has been modified by removal of the region encoding its export signal. Mutagenesis of Streptococcus pyogenes with TnFuZ ("FuZ" stands for fusions to phoZ) identified genes encoding secreted proteins whose expression was enhanced during growth in an aerobic environment. Thus, TnFuZ should be valuable for analysis of protein secretion, gene regulation, and virulence in gram-positive bacteria.

* Corresponding author. Mailing address: Department of Molecular Microbiology, Washington University School of Medicine at Washington University Medical Center, Campus Box 8230, 660 S. Euclid Ave., St. Louis, MO 63110-1093. Phone: (314) 362-1485. Fax: (314) 362-1232. E-mail: caparon{at}borcim.wustl.edu.

Applied and Environmental Microbiology, February 2002, p. 928-932, Vol. 68, No. 2
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.02.928-932.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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