人白细胞介素18的基因克隆及其在大肠杆菌中的表达

Cloning and Expression of Human IL-18 Gene in E.coli

目的 在大肠杆菌中高效表达IL 18。方法 从人外周血中提取RNA ,用RT -PCR方法得到IL 18的cDNA ,双酶切将其插入pET 2 3(b) + 载体中 ,转化大肠杆菌DH5α。筛选阳性菌落 ,提取质粒 ,转化大肠杆菌BL2 1(DE3)中进行表达。结果 所获得人IL 18克隆 ,经测序与GenBank报道的完全一致。所表达的产物经SDS -PAGE显示在相对分子质量 2 0 0 0 0处出现一条特殊条带 ,与预期的分子量一致。结论 已成功获得了人IL

Objective To highly express IL 18 in E.coli .Methods Extract RNA from human peripheral blood,amplify IL 18 cDNA by RT-PCR,insert into pET 23(b)+vector and transform to E.coli DH5 α.Select positive colonies,extreact recombinant plasmid and transform to E.coli BL21(DE3) for expression.Results The sequence of obtained human IL 18 clones was completely identical to that of IL-18 in GenBank.SDS PAGE proved that the relative molecular weight of the expressed product was about 20000,which was consistent with respected.Conclusion\ Human IL 18 gene was successfully cloned and expressed in E.coli.