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XcpT overproduction interferes with Xcp-dependent secretion. PAO1 strains containing pMTWT (xcpT gene cloned into pMMB190) where indicated or pMMB190 were grown on skim-milk plates in the presence of 2 mM IPTG where indicated. (A and B) A loop of cells was collected from plates and analyzed for XcpT and XcpP production by Western blotting with appropriate antibodies. (C) The zones of hydrolysis observed around the colonies are mostly due to the proteolytic activity of elastase (LasB).
