Fig. 3
Complex of purified KsgA, KsgA (1-213), or KsgA (9-273) with double-stranded oligonucleotides. Oligonucleotide substrates (10 fmol) were incubated at 4 °C for 60 min with purified proteins (100 pmol). Samples were separated by 12% PAGE. (a) Oligonucleotides used in the gel mobility shift assay. The asterisk indicates the base is present on the DNA strand labelled with 32P. The red letter means a damaged base or its replacement with a normal base, and the blue letter means a base that is a pair of the red letter base. Oligonucleotides contain (b) Tg/C (lanes 1-4) and T/A (lanes 5-8), (c) 5-foU/C (lanes 1-4), and (d) 5-ohC/G (lanes 1, 6, 11, and 16), 5-ohC/A (lanes 2, 7, 12, and 17), 5-ohC/T (lanes 3, 8, 13, and 18), 5-ohC/C (lanes 4, 9, 14, and 19), and G/C (lanes 5, 10, and 15). (b) Lanes 1 and 5, no protein; lanes 2 and 6, purified His-KsgA; lanes 3 and 7, purified His-KsgA (1-213); lanes 4 and 8, purified His-KsgA (9-273). (c) Lane 1, no protein; lane 2, purified His-KsgA; lane 3, purified His-KsgA (1-213); lane 4, purified His-KsgA (9-273). (d) Lanes 1-4, no protein; lanes 5-9; purified His-KsgA; lanes 10-14, purified His-KsgA (1-213); lanes 15-19, purified His-KsgA (9-273)