Deschamps J R, Miller C E, Ward K B
Naval Research Laboratory, Washington, DC 20375, USA.
Protein Expr Purif. 1995 Aug;6(4):555-8. doi: 10.1006/prep.1995.1073.
The green fluorescent protein (GFP) of the jelly fish Aequoria victoria was cloned into an Escherichia coli cell line that is a methionine auxotroph. The recombinant GFP (rGFP) was isolated from the cells and purified using a simple procedure consisting of only two chromatographic steps: size-exclusion chromatography and ion-exchange HPLC. Due to the hydrophobic nature of the protein, the surface characteristics of the HPLC size column, and the high initial salt concentration, the rGFP sticks to the size column and is eluted by reducing the salt concentration. Due to this unique behavior the purification procedure can readily be scaled to handle larger quantities of rGFP.
将维多利亚多管水母的绿色荧光蛋白（GFP）克隆到一种甲硫氨酸营养缺陷型的大肠杆菌细胞系中。从细胞中分离出重组绿色荧光蛋白（rGFP），并通过一个仅包含两个色谱步骤的简单程序进行纯化：尺寸排阻色谱和离子交换高效液相色谱。由于该蛋白质的疏水性、高效液相色谱尺寸柱的表面特性以及较高的初始盐浓度，rGFP会黏附在尺寸柱上，并通过降低盐浓度来洗脱。由于这种独特的行为，纯化程序可以很容易地扩大规模以处理更大数量的rGFP。
