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Figure 2 | BMC Molecular Biology

Figure 2

From: Biochemical characterisation of LigN, an NAD+-dependent DNA ligase from the halophilic euryarchaeon Haloferax volcanii that displays maximal in vitro activity at high salt concentrations

Figure 2

Salt requirements for LigN activity. A. DNA ligase activity assays performed under standard conditions (as described in the Methods) at increasing KCl concentrations, from 0 to 3.2 M in 400 mM steps (lanes 1–9). Quantitation of the data is shown below, with the product quantity obtained in 3.2 M KCl (lane 9) indicated as 100%. B. DNA ligase activity assays performed as described in Methods with LigN and 3.2 M KCl (lane 1), 3.2 M NaCl (lane 2), 3.2 M potassium acetate (lane 4), 3.2 M sodium acetate (lane 5) or with no added enzyme (lane 5, reaction performed in 3.2 M KCl). Quantitation of the data is shown below, with the product quantity obtained in 3.2 M KCl (lane 1) indicated as 100%. C. DNA ligase activity assays performed under standard conditions (see Methods) with no added enzyme (lane 1, reaction mix contains 10 mM magnesium acetate) or with LigN and no divalent cation (lane 2), 10 mM MgCl2 (lane 3), 10 mM MnCl2 (lane 4), 10 mM CaCl2 (lane 5), 10 mM CoCl2 (lane 6), 10 mM NiCl2 (lane 7) and 10 mM ZnCl2 (lane 8). Quantitation of the data is shown to the right, with the product quantity obtained in 10 mM MgCl2 (lane 3) indicated as 100%.

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