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DnaK-His was induced by 1 mM IPTG. Cells were harvested after 3 h, resuspended in Buffer C (50 mM Tris-HCl (pH 7.5), 10 (v/v) glycerol) containing 0.1 lysozyme, and kept on ice for 30 min. The cells were disrupted by sonication, and the resulting lysate was centrifuged at 75,000 g for 90 min. The supernatant was loaded onto a Ni2 -NTA-agarose (Qiagen) column equilibrated with Buffer C. Proteins
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