Religation Kinetics.

<p>(a) Gel analysis of the religation kinetics observed when incubating the hTop1- or hTop1(pf-Linker)-suicide cleavage complex (Cl1) with the R11 complementary ligator oligonucleotide (shown at the top of the figure) in absence [lanes 3-6 and lanes 12-15 for hTop1 and hTop1(pf-Linker) respectively] or in presence of 100 µM CPT [lanes 7-10 for hTop1 and lanes 16-19 for hTop1(pf-Linker)]. In lane 1 no protein was added. The lanes 2 and 11 represent the time 0 for hTop1 and hTop1(pf-Linker) mediated reactions, respectively. “Cl1” represents the DNA fragment cleaved at the preferred enzyme site; “religation” is the restored fully duplex oligonucleotide representing the final product of the religation reaction. (b) Plot of the percentage of religation normalized to the plateau value, in absence or in presence of CPT for the hTop1 (full and dashed black lines, respectively) and hTop1(pf-Linker) (full and dashed grey lines, respectively). Data shown are means ± SD from 3 independent experiments.