GST-EGF融合蛋白在大肠杆菌中的表达与纯化  

Expression of GST EGF Fusion Protein in E coli and Its Purification

作　　者：李文清[1] 徐柏年[1] 朱坚 罗进贤[1] 

机构地区：[1]中山大学生命科学学院

出　　处：《中山大学学报（自然科学版）》1998年第3期13-16,共4页Acta Scientiarum Naturalium Universitatis Sunyatseni

基　　金：广东省"八五"重点资助

摘　　要：将小鼠ＥＧＦ基因克隆至谷胱甘肽Ｓ－转移酶（ＧＳＴ）融合表达载体ｐＧＥＸ－２Ｔ，转化大肠杆菌ＤＨ５α，获得高效表达，融合蛋白ＧＳＴ－ｍＥＧＦ经Ｓｅｐｈａｒｏｓｅ４Ｂ－ＧＳＨ亲和层析柱纯化和凝血酶消化获得有免疫活性的ｍＥＧＦ。Epidermal growth factor (EGF) enhances the growth of various cell types and has wide clinical applications. EGF gene has been expressed in E coli but its expressed product is usually unstable because of its low molecular weight. In this study, mouse EGF gene was cloned into GST gene fusion vector pGEX 2T. The resultant plasmid pGEX 2T EGF was used to transform E coli DH5α,where the GST EGF fusion protein was expressed efficiently. After purification with Sepharose 4B GSH affinity column and digestion with thrombin, immunoactive mEGF was obtained, indicating that the GST fusion gene system is not only able to increase the gene product stability but also helpful to product purification.

关 键 词：GST-EGF 基因表达 大肠杆菌 融合蛋白 

分 类 号：Q51[生物学—生物化学] Q78