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Applied and Environmental Microbiology, November 2007, p. 7232-7239, Vol. 73, No. 22
0099-2240/07/$08.00+0     doi:10.1128/AEM.01030-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Evaluation of the Staphylococcus aureus Class C Nonspecific Acid Phosphatase (SapS) as a Reporter for Gene Expression and Protein Secretion in Gram-Negative and Gram-Positive Bacteria{triangledown}

Erika du Plessis,1* Jacques Theron,2 Eldie Berger,1 and Maureen Louw1

CSIR Biosciences, Box 395, Pretoria 0001, South Africa,1 Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria 0002, South Africa2

Received 9 May 2007/ Accepted 19 September 2007

A phosphatase secreted by Staphylococcus aureus strain 154 has previously been characterized and classified as a new member of the bacterial class C family of nonspecific acid phosphatases. As the acid phosphatase activity can be easily detected with a cost-effective plate screen assay, quantitatively measured by a simple enzyme assay, and detected by zymography, its potential use as a reporter system was investigated. The S. aureus acid phosphatase (sapS) gene has been cloned and expressed from its own regulatory sequences in Escherichia coli, Bacillus subtilis, and Bacillus halodurans. Transcriptional and translational fusions of the sapS gene with selected heterologous promoters and signal sequences were constructed and expressed in all three of the host strains. From the range of promoters evaluated, the strongest promoter for heterologous protein production in each of the host strains was identified, i.e., the E. coli lacZ promoter in E. coli, the B. halodurans alkaline protease promoter in B. subtilis, and the B. halodurans {sigma}D promoter in B. halodurans. This is the first report on the development of a class C acid phosphatase gene as a reporter gene with the advantage of being able to function in both gram-positive and gram-negative host strains.

* Corresponding author. Mailing address: CSIR Biosciences, Box 395, Pretoria 0001, South Africa. Phone: 27 12 841-2192. Fax: 27 12 841-3651. E-mail: emdplessis{at}csir.co.za

{triangledown} Published ahead of print on 28 September 2007.

Applied and Environmental Microbiology, November 2007, p. 7232-7239, Vol. 73, No. 22
0099-2240/07/$08.00+0     doi:10.1128/AEM.01030-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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