PRODUCTION OF THROMBIN INHIBITOR
PURPOSE:To efficiently produce desulfatohirudin protein, by culturing an Escherichia coli transformed with a plasmid into which the DNA coding the hirudin protein of thrombin-inhibiting activity is inserted. CONSTITUTION:A DNA sequence containing the DNA coding the hirudin protein which inhibits the...
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Sprache: | eng |
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Zusammenfassung: | PURPOSE:To efficiently produce desulfatohirudin protein, by culturing an Escherichia coli transformed with a plasmid into which the DNA coding the hirudin protein of thrombin-inhibiting activity is inserted. CONSTITUTION:A DNA sequence containing the DNA coding the hirudin protein which inhibits the thrombin is inserted into the manifestation vector to form a recombinant plasmid. The recombinant plasmid is used to transform Escherichia coli and the transformed product is cultured. The culture mixture is centrifuged to collect the cell bodies, then, the cell bodies are treated with ultrasonic waves in a buffer solution and lysozyme or dynomill to prepare the supernatant. The supernatant is fractionated using ammonium sulfate or subjected to gel filtration with Sephadex G-50 or the like to obtain the desulfatohirudin protein. |
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