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.
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.
du Plessis
Erika
E
CSIR Biosciences, Box 395, Pretoria 0001, South Africa. emdplessis@csir.co.za
Theron
Jacques
J
Berger
Eldie
E
Louw
Maureen
M
D000135:Acid Phosphatase / Q000235:genetics* / Q000378:metabolism; D001412:Bacillus subtilis / Q000235:genetics; D001431:Bacteriological Techniques / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter / Q000235:genetics*; D005821:Genetic Techniques; D006090:Gram-Negative Bacteria / Q000235:genetics*; D006094:Gram-Positive Bacteria / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D013211:Staphylococcus aureus / Q000201:enzymology* / Q000235:genetics
D011993:Recombinant Fusion Proteins; D000135:Acid Phosphatase