Skip to main content
Figure 3 | BMC Molecular Biology

Figure 3

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 3

Additional properties of LigN. A. LigN activity assays performed at 40°C, 45°C, 50°C and 55°C as indicated in the Methods except only 0.128 pmol of LigN protein (0.2 μg) was used per 20 μl assay. Quantitation of the data is shown to below, with the quantity of product obtained at 40°C indicated as 100%. B. Assays performed under standard conditions (45°C, as indicated in the Methods) following pre-incubation of the LigN protein for 30 minutes at 45°C, 55°C, 65°C or 75°C (lanes 2–5 respectively), or with no LigN protein added (lane 1). Quantitation of the data is shown below, with the quantity of product obtained following preincubation at 45°C indicated as 100%. C. LigN activity assays (20 μl) in the presence (lanes 1–5) or absence (lanes 6–10) of 1 mM NAD+ with 0.64 pmol LigN (lanes 1, 6), 0.128 pmol (lanes 2, 7), 25.6 fmol (lanes 3, 8), 5.12 fmol (lanes 4, 9) and 1.024 fmol (lanes 5, 10) per reaction. The reaction was incubated at 45°C for 6 hours. Quantitation of the data in lanes 3–5 (with NAD+) and 8–10 (no added NAD+) is shown below, with the product quantities obtained lanes 3 and 8 indicated as 100% for the appropriate datasets.

Back to article page

BMC Molecular Biology

ISSN: 1471-2199

Contact us