Affiliation:
1. CSIR Biosciences, Box 395, Pretoria 0001, South Africa
2. Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria 0002, South Africa
Abstract
ABSTRACT
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
σ
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.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
3 articles.
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