基因重组蛋白L243诱导表达包涵体的变性、复性与纯化

目的 对来自于日本七鳃鳗的基因重组蛋白L243包涵体进行变性、复性与纯化,以获得成分均一的活性蛋白.方法 以6 mol/L尿素为变性剂,以0.1 mol/L氧化型谷胱甘肽及0.9 mol/L还原型谷胱甘肽为复性剂,对构建于pET23b的日本七鳃鳗L243基因在大肠杆菌Rosetta中表达的包涵体进行了的变性、复性与组氨酸亲和层析纯化.结果 经变性、复性后,获得了均质的L243纯化蛋白,该蛋白的复性率达80%.结论 成功对上述重组蛋白包涵体进行了变性、复性及纯化,为后续与此蛋白相关的生物活性测定奠定了物质基础.

Denaturation, renaturation and purification of the inclusionbodies from recombinant L243 proteint

Objective To obtain the soluble homogeneous protein, Denaturation, renaturation and purification of the inclusionbodies from recombinant L243 was carried out. Methods The inclusionbodies of recombinant L243 proteint were expressed in E.coli of Rosetta, and it was purified by using the His-Bind Column Chromatography under 6 mol/L urea condition. The inclusionbodies were denatured under 6 mol/L urea, and its' renaturation were under 0.6 mol/L urea, 0.1 mol/L OxydizedGlutathione and 0.9 mol/L Reducedglutathione. Results After denaturation and renaturation, we obtained the soluble homogeneous protein with biological activities. The rate of renaturation was about 80%. Conclusion The denaturation and renaturation of above recombinant protein were successful, which laid the basis of studying the biological activities related to the recombinant L243 proteint.