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To purify UhpT, the sugar phosphate carrier ofEscherichia coli,we constructed a variant (HisUhpT) in which 10 tandem histidine residues were placed at the UhpT N terminus and then used Ni 2+ –agarose affinity chromatography of detergent-solubilized proteins. Membrane vesicles from a strain overexpressing HisUhpT were extracted at pH 7.4 with either 1.5%n-octyl-β-d-glucopyranoside (octylglucoside)...
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