Journal: Infection and Immunity
Article Title: Exposure of Salmonella enterica Serovar Typhimurium to a Protective Monoclonal IgA Triggers Exopolysaccharide Production via a Diguanylate Cyclase-Dependent Pathway
doi: 10.1128/IAI.00813-12
Figure Lengend Snippet: Expression of an EPS containing O-Ag capsule-like material by S. Typhimurium in response to Sal4. (A and B) EPS preparations from WT strain 14028 or strain SJF12 (ΔoafA) treated with Sal4 (10 μg/ml), 23D7 (10 μg/ml), or CDM for 4 h at 37°C were separated by SDS-PAGE on 12% gels with an extra-long (35-mm) stacking gel. (A) Silver stain. Lane L, molecular weight ladder; lane 1, WT treated with CDM; lane 2, WT treated with Sal4; lane 3, WT treated with 23D7; lane 4, heat-killed WT treated with Sal4; lane 5, SJF12 treated with CDM; and lane 6, SJF12 treated with Sal4. (B) Following SDS-PAGE, EPS preparations were transferred onto nitrocellulose membranes and subjected to Western blotting using Salmonella-specific O-Ag polyclonal antisera, as described in Materials and Methods. Lane L, ladder; lane 1, WT treated with CDM; lane 2, WT treated with Sal4. (C) Cellulose production in response to Sal4. WT and ΔyeaJ strains were spotted on CF agar plates and then overlaid with 5 μl Sal4 (47 μg/ml) or CDM. Plates were then incubated at 37°C for 24 h and photographed on a UV light box.
Article Snippet: Rabbit polyclonal antiserum against Salmonella O antigens (group B factors 1, 4, 5, and 12) was purchased from BD Difco (Franklin Lakes, NJ). . Construction of Δ yeaJ , Δ yeaJ (pYeaJ), bscE :: kan (pYeaJ), and YeaJ overexpression strains.
Techniques: Expressing, SDS Page, Silver Staining, Molecular Weight, Western Blot, Incubation