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SDS-PAGE analysis of Sarkosyl-insoluble OMPs of V. parahaemolyticus.

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posted on 2014-08-22, 03:34 authored by Tomotaka Tanabe, Ayaka Kato, Keiichi Shiuchi, Katsushiro Miyamoto, Hiroshi Tsujibo, Jun Maki, Shigeo Yamamoto, Tatsuya Funahashi

SDS-PAGE analysis was performed with VPD5, VPD107 (seven iron-repressible OMRs-deficient mutant derived from VPD5), VPD108 (peuA-deficient mutant derived from VPD107), VPD108/pRK415-peuA, VPD109 (peuRS-deficient mutant derived from VPD107), and VPD109/pRK415-peuRS. The OMP fractions were prepared from cells grown in LB-Tris medium at pH 7.0, LB-Tris/+EDDA media at pH 7.0 and 8.0, or LB-Tris/+EDDA/+Ent media at pH 7.0 and 8.0. Lanes 1–7 and 9–10 were loaded with 20 µg OMPs, and lane 8 was loaded with 3 µg OMPs. Electrophoresis was performed on 7.5% SDS-polyacrylamide gels (130 mm long) at a constant current of 15 mA at 4°C. The gel was stained with Coomassie Brilliant Blue. The figure shows only the relevant portions of the gel. The iron-repressible OMPs expressed by VPD5 at pH 7.0 under iron-limiting conditions are boxed in lane 2. Lane M, molecular weight marker proteins; closed arrowheads, PeuA.

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