Yakhnin A V, Vinokurov L M, Surin A K, Alakhov Y B
Branch of the Institute of Bioorganic Chemistry RAS, Pushchino, Moscow Region, 142292, Russia.
Protein Expr Purif. 1998 Dec;14(3):382-6. doi: 10.1006/prep.1998.0981.
Green fluorescent protein (GFP) is widely used as an excellent reporter molecule in biochemistry and cell biology. Some biochemical and immunological assays require high-purity GFP. However, the majority of current procedures for GFP purification include multiple time-consuming chromatography steps with a low yield of the desired product or require tag-containing proteins. An alternative method is described for the GFP purification without affinity extensions using organic extraction yielding a highly homogeneous protein indistinguishable in spectroscopic properties from that purified by previous methods.
绿色荧光蛋白(GFP)在生物化学和细胞生物学中被广泛用作优良的报告分子。一些生化和免疫分析需要高纯度的GFP。然而,目前大多数GFP纯化方法包括多个耗时的色谱步骤,所需产物的产率较低,或者需要含标签的蛋白质。本文描述了一种替代方法,即使用有机萃取法纯化GFP,无需亲和延伸,得到的蛋白质高度均一,其光谱性质与以前方法纯化的蛋白质无法区分。