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Protein purification using intein-hydrophobin tag and alcohol precipitation

机译:使用内含蛋白-疏水蛋白标签和乙醇沉淀纯化蛋白质

摘要

A method of downstream purification of a recombinant protein in fusion with a hydrophobin-intein tag using alcohol precipitation. The method comprises (1) constructing a plasmid expressing a fusion protein in a host cell, wherein the fusion protein includes a target protein domain, an intein, and a hydrophobin domain; (2) culturing the host cell transfected with the plasmid forming a cell culture medium; (3) separating a cell culture supernatant containing the fusion protein from the cell culture medium; and (4) purifying a substantial amount of the target protein from the cell culture supernatant without its hydrophobin-intein fusion tag using alcohol precipitation.
机译:一种使用醇沉淀与疏水蛋白-内含子标签融合的下游纯化重组蛋白的方法。该方法包括:(1)在宿主细胞中构建表达融合蛋白的质粒,其中所述融合蛋白包括靶蛋白结构域,内含肽和疏水蛋白结构域; (2)培养用质粒转染的宿主细胞,形成细胞培养基。 (3)从细胞培养基中分离含有融合蛋白的细胞培养上清液。 (4)使用酒精沉淀法从细胞培养上清液中纯化大量目标蛋白,而没有其疏水蛋白-内含蛋白融合标签。

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