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Lon preferentially degrades tmRNA-tagged λ-CI-N protein in vitro. In vitro proteolysis assays were carried out at 37°C in a minimal activity buffer containing 50 mM Tris-HCl (pH 8.0), 10 mM MgCl2, and 1 mM dithiothreitol. Complete reaction mixtures contained 1 μM Lon, 10 μM substrate, and an ATP regeneration system. Time point samples were taken at the indicated times and analyzed by Tris-Tricine-PAGE followed by Coomassie blue staining.
