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Fig. 5 | Mobile DNA

Fig. 5

From: Characterization of a relaxase belonging to the MOBT family, a widespread family in Firmicutes mediating the transfer of ICEs

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

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Mobile DNA

ISSN: 1759-8753