鸡红细胞组蛋白八聚体制备。

CSH protocols Pub Date : 2008-12-01 DOI:10.1101/pdb.prot5112
Craig L Peterson, Jeffrey C Hansen
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引用次数: 12

摘要

核心组蛋白可以从多种细胞来源中纯化,包括果蝇胚胎、HeLa组织培养细胞、小牛胸腺或鸡红细胞。鸡红细胞是细胞组蛋白的极好来源:大量的来源材料很容易获得,纯化的组蛋白具有低水平的翻译后修饰,并且连接蛋白也可以从相同的细胞样本中纯化。此外,鸟类组蛋白具有与人类组蛋白相同的氨基酸序列。组蛋白原液可在4°C条件下成功保存一年以上,在-20°C条件下可保存数年。用这种方法,200毫升血液通常产生超过50毫克的纯化组蛋白八聚体。另外还介绍了H1和H5连接体组蛋白的纯化,以及H3/H4四聚体和H2A/H2B二聚体的制备。
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Chicken erythrocyte histone octamer preparation.

INTRODUCTIONCore histones can be purified from a variety of cell sources, including Drosophila embryos, HeLa tissue culture cells, calf thymus, or chicken erythrocytes. Chick erythrocytes are an excellent source of cellular histones: Large quantities of source material are readily obtainable, the purified histones have low levels of post-translational modifications, and linker histones can also be purified from the same cell sample. Also, avian histones have an amino acid sequence identical to that of human histones. Histone stocks can be stored successfully for more than a year at 4°C and for several years at -20°C. With this protocol, 200 mL of blood usually yields in excess of 50 mg of purified histone octamers. Additional optional procedures are also presented for the purification of H1 and H5 linker histones, as well as for the preparation of H3/H4 tetramers and H2A/H2B dimers.

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