【摘要】 目的 :构建hIL 2 /mGM CSF原核表达载体 ,获得具有hIL 2和mGM CSF活性的hIL 2 /mGM CSF融合蛋白。方法 :利用PCR重叠延伸术 ,将人白细胞介素 2 (hIL 2 )和鼠粒细胞、巨噬细胞集落刺激因子 (mGM CSF)基因 ,通过 2个脯氨酸 (Pro)基因的中间接头连接起来 ,扩增出hIL 2 /mGM CSF融合基因 ,并克隆于表达载体pBV2 2 0和pLY4中。结果 :获得序列正确的hIL 2 /mGM CSF融合基因 ,并在大肠杆菌中成功表达 ,表达率在 2 0 %左右。活性测定其hIL 2活性为 4 5×10 5U/mg ,mGM CSF活性为 3 85× 10 6U/mg。结论 :获得了具有hIL 2和mGM CSF活性的hIL 2 /mGM CSF融合蛋白 ,为研究hIL 2 /mGM CSF融合蛋白的新生物学功能奠定基础。更多还原

【Abstract】 Objective:hIL 2/mGM CSF fusion gene was constructed and expressed in E.coli.recombinant hIL 2/mGM CSF fusion protein had both biological activity of hIL 2 and mGM CSF.Methods:The hIL 2 and mGM CSF genes were amplified by PCR (splicing by overlap extension,SOE) and ligated with two prolines linkers.Then the fusion gene was cloned into vector pLY4 and PBV220.The recombinant plasmid were transfected into E.coli and expressed.Results:The sequence of hIL 2/mGM CSF fusion gene was correct the hIL 2/mGM CSF fusion protein was highly expressed in E.coli and that comprises 20% of total bacterial protein.The results of biological activity assay showed that the expressed product possessed both of the activities of hIL 2(4.5×10 5 U/mg) and mGM CSF(3.85×10 6 U/mg).Conclusion:Have successful constructed the recombinant hIL 2/mGM CSF fusion protein which possessed bioactivities similar to that of the natural hIL 2 and mGM CSF.更多还原