hIL-1Ra基因的克隆及在原核细胞中的表达

Cloning of hIL-1Ra gene and its expression in E.coli

目的 :克隆hIL 1Ra基因并在大肠杆菌中高效表达。方法 :从人外周血中分离白细胞并提取其总RNA。用RT PCR获得hIL 1RacDNA ,克隆入pBV2 2 0表达载体进行诱导表达。对表达产物进行复性和纯化。结果 :成功地构建了pBV2 2 0 IL 1Ra表达载体 ,并在大肠杆菌中获得高效表达 ,表达量占全菌的 4 0 %。对表达产物进行分离纯化及活性分析表明 ,获得纯度大于98%的样品。该样品具有明显抑制IL 1刺激EL 4细胞分泌IL 2的作用。结论 :在大肠杆菌中成功地表达有活性的hIL 1Ra基因 ,为进一步的开发利用奠定了实验基础。

AIM: To clone human IL-1Ra gene and express it in E.coli. METHODS: Human IL-1Ra cDNA was obtained by RT-PCR with the the total RNA extracted from human peripheral blood leucocytes as template. The cDNA was cloned into pBV220 vector and expressed. The expressed product was renatured and purified. Separation, purification and bioactivity analysis of the expressed products were performed. RESULTS: IL-1Ra gene was successfully expressed in E.coli and the expression level reached to about 40% of total bacteria protein. The purity of the final product was over 98%. The product could obviously suppress the secretion of IL-2 by EL-4 cells stimulated with IL-1β. CONCLUSION: The expression of hIL-1Ra gene with bioactivity in E.coli lays experimental foundation for further development and utilisation.