FIG 7.

Influence of the lack of five TFs on the expression of Rsd protein. (A) The wild type and five mutants, each lacking the indicated gene on top, were grown in medium E containing 2% polypeptone and 0.5% glucose. Cells were harvested in the early stationary phase. The total amount of Rsd in whole-cell extract was measured by Western blotting, while the amount of affinity-purified His-tagged RpoD was measured by protein staining with Coomassie brilliant blue (CBB). The level of Rsd protein, shown in panel C, was determined by densitometry of the gel pattern and is shown as the relative value to that in wild-type cells. (B) His-tagged RpoD-Rsd complexes were affinity purified. The amount of Rsd bound on this complex was measured by immunoblotting. The total amount of His-tagged RpoD was observed by protein staining with CBB. The accuracy of immunoblot measurement by the method herein employed is more than 90% (56).