Fig 6.
Pel-reactive material is not detected in the periplasmic fraction. (A) PAO1ΔwspFΔpsl PBADpel, the pelA catalytic mutant D528A, and PAO1 expressing pPSV18 were assessed for Pel production by dot blotting. PAO1ΔwspFΔpsl PBADpel strains were grown in the presence or absence of 0.5% arabinose as indicated. Pel polysaccharide expression was analyzed for the EDTA-extracted whole-cell pellet, periplasmic fraction, and EDTA-extracted spheroplasts. (B) The whole-cell (WC) fraction, EDTA-extracted whole-cell pellet, periplasmic fraction, and EDTA-extracted spheroplasts were probed for PelC (membrane protein), RNA polymerase (RNAp; cytosolic protein), and β-lactamase (Bla; periplasmic protein).