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. 2019 May 3;10:18. doi: 10.1186/s13100-019-0160-9

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© The Author(s). 2019

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 5 — RelSt3 harbours nicking activity. In all assays, 200 ng of pBR322-oriT plasmidic DNA were incubated with the indicated amount of RelSt3 protein. a RelSt3 nicking activity according to temperature. pBR322-oriT plasmid was incubated without or with 2 μg of RelSt3 protein for 20 min at 4, 10, 20, or 30 °C in the presence of 5 mM Mn²⁺. FI: supercoiled plasmid DNA. FII: open circular form. FIII: linear form. b Time course in the presence of 2 μg of RelSt3 and 5 mM Mn²⁺ at 30 °C. The nicking activity was established by band quantification monitoring the decrease of FI form. c Dose response of RelSt3 nicking activity. DNA was incubated for 20 min at 30 °C with 5 mM Mn²⁺ without (lane 1) or with 0.02 (lane 2), 0.05 (lane 3), 0.2 (lane 4), 0.5 (lane 5), 1 (lane 6), 2 (lane 7), 5 (lane 8), 7 (lane 9) or 10 (lane 10) μg of RelSt3 protein