摘要
目的：根据胰岛素样生长因子１（ｉｎｓｕｌｉｎｌｉｋｅｇｒｏｗｔｈｆａｃｔｏｒ１，ＩＧＦ１）的三维结构及其与ＩＧＦ结合蛋白３（ＩＧＦＢＰ３）的结合位点，构建ＩＧＦ１突变体。方法：以合成的人ＩＧＦ１基因为模板，利用体外定点突变技术，获得［Ｔｙｒ１５，Ｌｅｕ１６］ＩＧＦ１突变基因，在大肠杆菌中进行表达，重组蛋白经纯化、复性后，还采用蛋白免疫印迹分析、荧光光谱分析和生物活性测定进行验证。结果与结论：成功地构建并表达了ＩＧＦ１突变体，［Ｔｙｒ１５，Ｌｅｕ１６］ＩＧＦ１的生物学活性并未受到影响。
Objective: To construct insulinlike growth factor 1 mutant(mIGF1) according to the three dimensional structure of IGF1 and its binding sites with IGF1 binding protein 3 Methods: mIGF1 of IGF1 was obtained with altered sites X]in vitro mutagenesis system using synthesized human IGF1 gene as the template, and expressed in E.coli . The purification and renaturation of mIGF1 were investigated. The recombinant mutant was characterized through Western blot, fluorescent chromatography and biological assay. Results and Conclusion: mIGF1 was successfully expressed in E.coli . The change from Q15, F16 to Y15, L16 had no effect on its biological activity.
出处
《军事医学科学院院刊》
CSCD
北大核心
1999年第2期94-96,共3页
Bulletin of the Academy of Military Medical Sciences
