FIG. 5.
Western blot analysis of PrfA and PrfA-regulated virulence proteins LLO and ActA. WT EGD-e (A and B), insertion mutants (::ccpA, ::hprK, and ::ptsH) and strain P14-A (A), and revertants (RccpA, RhprK, and RptsH) (B) were grown in BHI medium. At an OD600 of 1.0 (later growth phase) proteins were prepared, equal amounts of proteins were separated by SDS-PAGE, and equivalent loading of the gels was controlled by Coomassie-staining (data not shown). Loading controls: PrfA, purified PrfA protein; ΔpkP-1, EGD with a deletion in the virulence gene cluster (18).