Figure 6.

Effect of inter-site distance on cleavage of head-to-head substrates. (A) Plasmid substrates used in the assays. Arrowheads denote the position and orientation of the 6 bp LlaGI target sequence. (B) Example of LlaGI sequences for n = 0 to 5 bp. Plasmid DNA (2 nM) was pre-incubated with 200 nM LlaGI for 2 min at 25°C. Cleavage was initiated by addition of ATP (to 4 mM). Samples were taken at the times indicated, stopped, and analyzed by agarose gel electrophoresis. The graphs show the amount of supercoiled substrate DNA remaining (C) and linear DNA produced (D) as a function of time following addition of ATP. Points are the average of two repeat experiments.