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In vitro binding assays using recombinant proteins Saturating amounts of clarified bacterial lysates containing GST and GST-S fusions were added to Glutathione Sepharose beads that were previously washed with lysis buffer (50 mM Tris, pH 7.4, 150 mM NaCl, 1 mM EDTA, 5 mM 2-mercaptoethanol, 1% Triton X-100) and incubated at 4 C for 1 h on a tube roller