一种“高性价比”切胶纯化原核表达蛋白的方法

目的：建立一种低成本高效率的纯化表达蛋白的新方法。方法：在保证不改变原有蛋白生物活性的前提下，以NaAc染色切胶纯化方法为参考，通过以KCl取代NaAc染色目的蛋白、反复冻融后快速离心等操作步骤取代原来的电泳后过夜透析等实验步骤。结果：最终使原实验时间至少缩短2/3，而且所得目的蛋白不存在浓度和纯度降低的问题。结论：为许多条件有限的实验室进行纯化原核表达蛋白的研究提供了一种“性价比”更高的实用方法。

A “Cost-effective” Method for Purification of Prokaryotic Expression Proteins in Gel Slices

To establish a new cost-effective method for purification of the recombinant protein,recombinant proteins expressed in E.coli were separated by using SDS-PAGE.Target proteins were isolated by cutting the gel slices that contained the right bands which were stained by 0.25 mol/L KCl instead of 4 mol/L NaAc solution.The author selected the modified experimental procedures of including repeated freezing and thawing then high-speed centrifugation to replace that of the original overnight dialysis after dialysis bag-electrophoresis elution.The author improved the experimental efficiency significantly.Proteins purified by this modified method remained proper antigenicity when detected by ELISA and western blot.Compared with other conventional ways of protein purification,this method was relatively simple and effective,especially suitable for the general laboratory.