Figure 5
From: Binding of the baculovirus very late expression factor 1 (VLF-1) to different DNA structures
Binding of *VLF-1 to hr-containing DNA analyzed by electrophoretic mobility shift assay. Reaction mixtures (10 μl) containing 0.004 pmol 32P-labeled oligonucleotide probe and *VLF-1 were incubated for 20 min at 23°C, fixed with 0.5% glutaraldehyde, and analyzed by electrophoresis in a native 6% polyacrylamide gel as described in Materials and Methods. (A) Reactions shown in lanes 1 to 3 contained linear double-stranded hr DNA (substrate VI), those in lanes 4–6 contained a mixture of hairpin hr DNA (substrate VII) and H-shaped (double Y-form) hr DNA (substrate VIII). *VLF-1 was added to the reaction mixtures in the following amounts: lanes 2 and 5, 25 ng; lanes 3 and 6, 50 ng. No protein was added to the reactions shown in lanes 1 and 4. No MgCl2 (B) Reactions shown in lanes contained the following DNA probes: lanes 1 to 6, a mixture of hairpin hr DNA (substrate VII) and H-shaped (double Y-form) hr DNA (substrate VIII); lanes 7 and 8, cruciform DNA lacking hr sequence (substrate V). *VLF-1 (35 ng) was added to the reactions shown in lanes 2 to 6, and 8. No protein was added to the reactions shown in lanes 1 and 7. The final concentrations of MgCl2 in the reaction mixtures were 1 mM (lane 3), 2 mM (lane 4), 5 mM (lane 5), and 10 mM (lane 6). No MgCl2 was added to the reactions shown in lanes 1, 2, 7, and 8.