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目的 :纯化在大肠杆菌中表达的重组人胰岛素样生长因子Ⅰ (rhIGF Ⅰ ) ,并对其性质进行鉴定。方法 :发酵表达菌株 ,提取包涵体 ,采用离子交换和凝胶过滤两步纯化 ,从相对分子质量、免疫学性质、荧光光谱、N端 15个氨基酸的序列分析等方面进行鉴定。另外 ,对重组蛋白进行了复性和生物学功能的检测。结果与结论 :纯化后rhIGF Ⅰ纯度可达 99%以上 ,相对分子质量、免疫印迹、荧光光谱及N端 15个氨基酸的分析结果与预计的相同 ,rhIGF Ⅰ促细胞生长的ED50 为 3~ 10ng/ml。
Objective: To purify recombinant human insulin - like growth factor Ⅰ (rhIGF Ⅰ) expressed in E. coli and identify its properties. Methods: The expressed strain was fermented and the inclusion bodies were extracted. The two strains were purified by ion exchange and gel filtration and identified from the relative molecular mass, immunological properties, fluorescence spectra and sequence analysis of 15 amino acids at the N terminus. In addition, the recombinant protein was tested for its refolding and biological functions. RESULTS AND CONCLUSION: The purity of rhIGF Ⅰ was over 99%. The relative molecular mass, immunoblot, fluorescence spectra and 15 amino acids of N terminal were the same as expected. The ED50 of rhIGF Ⅰ cell growth was 3 ~ 10ng / ml.