FIG. 1.

Purification of AdpA-H (A) and binding of AdpA-H to the upstream activation sequence of strR (B). (A) Appropriate amounts of the insoluble (lane 1) and soluble (lane 2) fractions prepared from E. coli cells harboring pET-adpA and about 0.2 μg of protein of the sample (lane 3) purified with His-bind resin were run. (B) A ³²P-labeled 100-bp fragment, including the AdpA-binding site upstream of strR, was prepared by PCR and used in the gel mobility shift assay. The positions of AdpA-H-bound (solid triangle) and free (open triangle) probes are shown. The position of the gel well is also indicated by an arrow. The retarded signals become stronger with an increase in the amount of AdpA-H. The amounts of AdpA-H were 0.04 μg (lane 2), 0.2 μg (lane 3), and 1 μg (lane 4). Lane 1 is a control lane in which no AdpA-H was contained.