FIG. 2.

Representative gel mobility shift assay used to establish the minimal sequence requirement for T-ag-obd_131–260 binding to site II. Lanes: 2 and 3, products of band shift assays conducted with the 64-bp core oligonucleotide and 3 or 6 pmol, respectively, of T-ag-obd_131–260; 5 and 6, products of band shift assays conducted with the 31-bp site II oligonucleotide and 3 or 6 pmol, respectively, of T-ag-obd_131–260; 8 and 9, products of band shift assays conducted with the 23-bp site II oligonucleotide and 3 or 6 pmol, respectively, of T-ag-obd_131–260. As a control for nonspecific binding, band shift assays were conducted with the 31-bp site IIm control oligonucleotide and either 3 or 6 pmol of T-ag-obd_131–260 (lanes 11 and 12). Lanes 1, 4, 7, and 10 contain the products of band shift assays conducted in the absence of protein with the indicated oligonucleotides. The input or free duplex DNA (F) is indicated by the arrow. The protein-to-oligonucleotide ratios with 3 and 6 pmol of T-ag-obd_131–260 and 25 fmol of oligonucleotide are 120:1 and 240:1, respectively.