FIG. 2.

Production and cellular localization of entire and truncated HasR polypeptides. C600 cells carrying pBAD24-hasR, pBAD24-hasR Δ11-91, pBAD24-hasR Δ11-192, or pBAD24-hasR 1-242 were grown in LB medium with and without 0.02% arabinose at 37°C to an OD₆₀₀ of 1. Total cell extracts and outer membrane and soluble fractions were prepared. A: Outer membrane and soluble fractions; 0.02 OD₆₀₀ equivalent of outer membrane fractions or 0.1 OD₆₀₀ equivalent of soluble fraction was loaded in each lane of an SDS-10% PAGE gel. HasR was immunodetected with a polyclonal anti-HasR serum used at a dilution of 1/5,000. B: Coomassie blue staining of total cell extracts and outer membrane fractions of arabinose-induced cells. Total cellular proteins precipitated with trichloroacetic acid and outer membrane fractons. For total cell extract analysis, 0.2 OD₆₀₀ equivalent of strains expressing wild-type HasR and HasR Δ11-91 and 0.4 OD₆₀₀ equivalent of the strain expressing HasR Δ11-192 were loaded. For outer membrane fractions, 1 OD₆₀₀ equivalent of strains expressing wild-type HasR and HasR Δ11-91 and 2 OD₆₀₀ equivalent of the strain expressing HasR Δ11-192 were loaded on the gels.