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A transfectant HeLa cell clone expressing HFE under the control of a tetracycline-repressible promoter was generated. HFE expression was fully repressed by the presence of doxycycline, while it was strongly induced by growth in the absence of doxycycline. HFE accumulation was accompanied by a large (~10-fold) decrease in H- and L-ferritin levels, by a ~3-4-fold increase in transferrin receptor, and...
The cDNA for streptavidin (residues 15–159) was subcloned into an expression vector in fusion at the N-terminus with the T7-tag (12 residues). Conditions were found to express the protein inEscherichia coliin a soluble, assembled, and active form. The protein was purified in two simple steps which involved heating at 75°C and affinity chromatography on iminobiotin agarose. The purified protein was...
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