Abstract
Gaussia luciferase (GLuc) is a secreted protein with significant potential for use as a reporter of gene expression in bacterial pathogenicity studies. To date there are relatively few examples of its use in bacteriology. In this study we show that GLuc can be functionally expressed in the human pathogen Staphylococcus aureus and furthermore show that it can be used as a biosensor for the agr quorum sensing (QS) system which employs autoinducing peptides to control virulence. GLuc was linked to the P3 promoter of the S. aureus agr operon. Biosensor strains were validated by evaluation of chemical agent-mediated activation and inhibition of agr. Use of GLuc enabled quantitative assessment of agr activity. This demonstrates the utility of Gaussia luciferase for in vitro monitoring of agr activation and inhibition.
Highlights
Staphylococcus aureus has been identified by the World Health Organization as a high priority pathogen [1]
Staphylococcal strains were cultured in tryptone soy broth (TSB) (Oxoid) and E. coli in lysogeny broth (LB) (Oxoid) with antibiotics added as required
Induction of agr with Respect to Growth Phase In S. aureus, agr-dependent quorum sensing is activated at high cell population densities when the autoinducing autoinducing peptide (AIP) reaches a threshold level
Summary
Staphylococcus aureus has been identified by the World Health Organization as a high priority pathogen [1]. It is one of the most common causes of bacterial infections in both hospitals and in the community [2]. As well as traditional drugs, alternative therapies are being investigated which are less likely to drive the development of resistance [5]. These include compounds which will target functions essential for infection such as colonization and virulence factor production without affecting bacterial growth [6]
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