恒波长同步荧光法测定吐温80-硫酸铵液固萃取体系中绿色荧光蛋白标记的融合蛋白

培养了含六聚组氨酸绿色荧光蛋白修饰融合蛋白的工程菌,对从工程菌中收集的蛋白原液用普通荧光和恒波长同步荧光2种荧光表征方式进行了比较,探究了分离蛋白的液固萃取体系中各因素(如硫酸铵浓度、吐温80浓度、酸度)对蛋白质荧光特性的影响,确定了萃取体系中适用的恒波长同步荧光测定法.运用该法得到液相的工作曲线线性相关系数达0.999 35,固相工作曲线线性相关系数为0.997 48,原始吐温80-硫酸铵液固萃取体系分离菌中目标蛋白,一次萃取固相收得率可达41.31%,调节pH 6.2时一次萃取固相收得率可达63.24%,进一步提高选择性,加入聚乙二醇修饰物可提高至93.50%.

Determination of Fusion Protein Tagged Green Fluorescent Protein in Tween80-Ammonium Sulphate Liquid-Solid Extraction System by Constant-Wavelength Synchronous Fluorescence

The engineering bacterium consisting of six-polyhistidine and green fluorescent protein(GFP) was cultivated.The general fluorescence and constant-wavelength synchronous fluorescence for the characterization of the protein obtained from the engineering bacterium were compared.The various factors involved in the liquid-solid extraction system such as the ammonium sulphate concentration,Tween 80 concentration and acidity on the characteristics of the fluourescence were explored.The method of constant-wavelengt...