FIG. 5.

Effect of a dsbA mutation on secretion of SPI1 TTS effector proteins. The strains contained the protein fusions indicated above the lanes and were either wild type or were deleted for hilC-D (which removed hilC-prgHIJK-hilD), dsbA, or cpxR. The SopA-M45 and SopB-M45 strains also contained the ara623::Tn10dTc insertion. Stationary-phase cultures were subcultured in either high-salt LB medium containing ampicillin and 0.2% l-arabinose (SopA-M45 and SopB-M45) or high-salt LB medium containing kanamycin (SlrP-CyaA) and were grown statically overnight. Culture supernatants were prepared as described Materials and Methods. Equivalent amounts of samples from the supernatants or whole-cell extracts of the strains were separated by SDS-12.5% PAGE. The resulting gels were blotted, and proteins were detected by using mouse anti-M45 (SopA-M45 and SopB-M45) or mouse anti-CyaA (SlrP-CyaA) and HRP-labeled rabbit anti-mouse secondary antibody. The strains used were JS356 through JS359 and JS364 through JS371. WT, wild type.