Abstract
A plasmid analogous to the one described by Nagai and Thogersen (Nature,309, 810–812, 1984) has been constructed for the expression of globins inE. coli. Induction with nalidixic acid produces high yields of a fusion protein, NS1-FX-β-globin, where NS1 represents 81 residues of a flu virus protein and FX represents a blood-clotting Factor Xa recognition sequence, Ile-Glu-Gly-Arg. This fusion protein is readily solubilized in 50 mM NaOH and remains in solution when thepH is adjusted to 8.6. Under these conditions, the fusion protein is hydrolyzed by activated Factor X, giving authentic β-globin which can be folded in the presence of cyanohemin and native α-chains to produce a tetrameric hemoglobin with the functional properties of natural human hemoglobin.
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Fronticelli, C., O'Donnell, J.K. & Brinigar, W.S. Recombinant human hemoglobin: Expression and refolding of β-globin fromEscherichia coli. J Protein Chem 10, 495–501 (1991). https://doi.org/10.1007/BF01025477
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DOI: https://doi.org/10.1007/BF01025477