Figure 2.
YsxC requirement for S. aureus growth. A) Strain LC109 (SH1000 Pspac~ysxC/pGL485) was grown on BHI agar plates containing 20 μg ml-1 Cam and 500 μM, 10 μM, 5 μM or 0 μM IPTG overnight. B) Exponentially growing cultures of strains SH1000 (●) and LC109 (SH1000 Pspac~ysxC/pGL485) (○,τ,ρ) were washed and resubcultured to approximately 1×106 CFU ml-1 in BHI (●) or in BHI supplemented with 20 μg ml-1 Cam plus different concentration of IPTG: 0 (○), 10 μM (▼) or 1 mM (△). Growth was monitored as CFU/ml. c) Western blot using anti-YsxC polyclonal antibodies. Strains SH1000 and LC109 (SH1000 Pspac~ysxC/pGL485) were grown to an OD600 = 0.5 in BHI and BHI plus 20 μg ml-1 Cam, respectively. Cells were harvested by centrifugation, the membrane protein fraction extracted and samples were separated by 12% (w/v) SDS-PAGE. Lane: 1, the size of molecular weight markers separated on the same gel is indicated; 2, SH1000; 3, LC109.