1887

Abstract

The glucosyltransferases of are recognized as important virulence factors for this cariogenic bacterium. To study the expression of the gene of in biofilms, a promoter ()–green fluorescent protein (GFP) reporter system was developed. A shuttle vector harbouring a :: cassette was introduced into GS-5, and the expression of GFP by the transformed cells was confirmed by fluorescence microscopy. Furthermore, confocal laser scanning microscopy was carried out on biofilms attached to polystyrene plates; enhanced expression was observed in various microcolonies across these biofilms. To further test the hypothesis that expression is upregulated in biofilms, flow cytometry analysis was done on planktonic and biofilm cells; this analysis showed an approximately five-fold increase in expression in the biofilm cells relative to the planktonic cells. Real-time (TaqMan) PCR analysis confirmed that expression in the biofilm cells was enhanced relative to the planktonic cells. Previously, it has been suggested that the gene might be co-transcribed with . Therefore, RT-PCR analysis was performed on -expressing ; this analysis demonstrated that was co-transcribed with . These results indicated that GFP expression can be utilized to examine gene regulation in biofilm formation.

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2002-11-01
2024-03-29
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