Figure 3
DNA ligase activity of Bsp-lig. Measured by molecular beacon assay (A-D) or urea-PAGE (E–G). (A) Specific activity of Bsp-lig (green circle) relative to Ame-lig (blue square) and T4 DNA ligase (purple triangle). (B) Relative activity of Bsp-lig with MgCl2 as the divalent cation (green circle) or MnCl2 (purple square). (C) Effect of NaCl on Bsp-lig activity). (D) Effect of ATP concentration on Bsp-Lig activity. (E) Bsp-lig activity with nicked substrate (Bsp-lig concentrations 20 nM, 10 nM, 5.0 nM, 2.5 nM, 1.8 nM). (F) Bsp-lig activity with cohesive-end substrate (Bsp-lig concentrations 550 nM, 55 nM, 20 nM, 10 nM). (G) Bsp-lig activity with mismatch substrate (Bsp-lig concentrations 550 nM, 55 nM, 20 nM, 10 nM). Uncropped gels are shown in Supplementary Fig. 7.
