| 摘要 |
PURPOSE: To obtain various brain-derived neurotrophic factors for a medicine, a diagnostic drug and a reagent, etc., having respectively different biological activity by centrifuging a host colibacillus-crushing solution producing a matured drain-derived neurotrophic factor, dividing into a supernatant and a guanidine hydrochloride-extracted solution of a precipitate, and purifying. CONSTITUTION: A gene-recombining colibacillus introduced of a recombining plasmid containing a gene coding for a matured brain-derived neurotrophic factor connected with a signal peptide gene ofβ-lactamase is cultured in a medium at 37 deg.C for 24 hours, centrifuged and resultant fungi are gathered. The fungi are suspended in a phosphoric acid buffer solution and crushed in an ultrasonic crusher, and resultant solution of crushed fungus is centrifuged to divide into a supernatant and a precipitate. The centrifuged precipitate is extracted with a guanidine hydrochloride aqueous solution to divide into a centrifuged supernatant and a guanidine hydrochloride-extracted solution of the centrifuged precipitate. These solutions are purified by an ion-exchange chromatography, a gel filtration or a reverse phase chromatography to obtain the objective brain-derived neurotrophic factors such as a medicine, a diagnostic drug and a research reagent, etc., having respectively different biological activity. |
