Figure 5.

Modulation of PolB1-mediated strand displacement by Sso7d in the presence of PCNA and RFC. (A) PolB1-mediated DNA strand displacement. PCNA (100 nM) and RFC (100 nM) were preincubated for 5 min at 70°C with P36/C72. PolB1 (5 nM) and various amounts of Sso7d were added. After 15 min at 70°C, the mixture was treated with proteinase K and extracted with phenol/chloroform. Reaction products were subjected to electrophoresis in 8% polyacrylamide gel containing 7 M urea in 1× TBE. Lanes 5–10, Sso7d concentrations were 0.5, 2.5, 10, 25, 50, 90 μM, respectively. (B) PolB1-mediated RNA strand displacement. Reactions were assembled and processed as described in (A) except that P36(5′RNA)/C72, instead of P36/C72, was used as the primer template.