Figure 4From: Genetic characterization of psp encoding the DING protein in Pseudomonas fluorescens SBW25Immunochemical analysis of Psp in P. fluorescens SBW25 and derived mutants. (A) Double immunodiffusion of cell supernatants and lysates. The central well in a 1.5% agarose plate contained rabbit anti-DING (Psp from SBW25) antiserum. The peripheral wells contained (clockwise, from the top) PBR826 (Δpsp) supernatant (Δpsp-sup), PBR828 (ΔhxcR) supernatant (ΔhxcR-sup), Psp standard (DING), SBW25 lysate (wt-lysate), PBR826 (Δpsp) lysate (Δpsp-lysate), PBR828 (ΔhxcR) lysate (ΔhxcR-lysate), SBW25 supernatant (wt-sup). (B) Western blotting of supernatants for SBW25 grown in different media. Concentrated cell supernatants were separated on a 12% SDS-PAGE gel, and Psp was detected with anti-human DING antiserum. Lane A, protein ladder; B, PBR826 (Δpsp) in LB broth (negative control); C, SBW25 in PL broth; D, E, SBW25 in PR broth; F, SBW25 in LB broth; G, recombinant Psp (positive control). (C) Western blotting of cell supernatants for SBW25 mutants grown on PL medium. Concentrated cell fractions were separated on a 12% SDS-PAGE gel, and Psp was detected with anti-Psp antiserum. Lane A and D, wild-type; B and E, PBR826 (Δpsp); C and F, PBR828 (ΔhxcR); G, Psp standard.Back to article page