FIG. 8.

Ugi does not inhibit Pa-MIG activity. Products of the glycosylase activity of 25 ng of Pa-MIG on a 96-bp heteroduplex DNA containing a U/G or T/G mismatch or a 5′-end-labeled 96-mer single-stranded DNA (20 fmol) containing U in the presence of Ugi were analyzed. One unit of E. coli UDG (Ec-UDG) or 25 ng of 70°C heat-treated Pa-MIG was incubated with Ugi (0, 3, and 10 U) at 37°C for 10 min before addition of the DNA substrates. After incubation at 50°C for 15 min, samples were treated with 4 μl of 1 M NaOH at 90°C for 4 min to cleave DNA at AP sites before electrophoresis. The uncut 96-mer DNA substrate (S) and cleaved 60-mer product (P) are indicated on the left. Asterisks indicate the 5′-end-labeled strand.