表皮生长因子与假单胞菌外毒素融合蛋白在大肠杆菌中的表达研究

Expression of recombinant fusion protein of human EGF with PE40 in E.coli

目的和方法：应用基因工程技术，将ＥＧＦｃＤＮＡ克隆到ｐＬＹ５／ＩＬ２－ＰＥ４０载体的ＥｃｏＲⅠ、ＳｍａⅠ位点之间ＩＬ２基因位置上，再将重组融合基因亚克隆到表达载体ｐＢＶ２２０的ＥｃｏＲⅠ、ＰｓｔⅠ位点上，以探索ＥＧＦ－ＰＥ４０融合基因在大肠杆菌ＤＨ５ａ中的进行表达及其生理功能。结果：ＳＤＳ－聚丙烯酰胺凝胶电泳和Ｗｅｓｔｅｒｎｂｌｏｔ表明：ＥＧＦ－ＰＥ４０融合蛋白获得表达，并且具有ＥＧＦ和ＰＥ４０的免疫学活性。

Abstract AIM and METHOD:In order to study the expression of fusion protein EGF-PE40 in E. coli DH 5a and the function, human EGF cDNA was cloned into the vector pLY5/IL2-PE40 at the EcoRⅠ and SamⅠ sites, where the IL2 gene was substituted. Subsequently recombined EGF-PE40 gene was subcloned into the expression Vector pBV220 of the EcoRⅠ and PstⅠ sites. RESULTS: A recombinant fusion protein was expressed successfully in high effect in E. coli confirmed by SDS-PAGE and Western blot. CONCLUSION:These results provide an important basis for EGF-PE40 protein and biological activities of the targetting tumor therapy. MeSH Eqidermal growth; Recombinant fusion proteins;