TNFHis-IL-11温度诱导融合表达载体的构建

目的构建TNFHis IL 11温度诱导融合表达载体。方法用EcoRI和PstI消化pGEMIL 11质粒后 ,回收IL 11基因片段 ,以同样的酶切位点克隆入TNFHis表达载体。将重组质粒转入大肠杆菌DH5α菌株 ,经 4 2℃温度诱导 4 .5h后做SDS PAGE和免疫活性鉴定。结果融合蛋白获得了表达 ,其分子量为 37kD ,其表达量占菌体总蛋白的 2 5 %。免疫印迹反应结果显示 ,表达的融合蛋白可与抗IL 11多克隆抗体产生阳性反应。该融合蛋白经盐酸羟胺裂解后可获得IL 11单体。结论TNFHis IL 11温度诱导融合表达载体构建成功。

Construction of TNFHis-IL-11 fusion expression vector

PurposeTo construct a temerature fusion protein expression vector(TNFHis IL 11).Methods After pGEMIL 11 plasmid was digested with EcoRI and PstI ,IL 11(550bp) gene was recovered and cloned into the sites of Eco RI and Pst I of TNFHis expression vector.The recombinant bacteria were induced at 42℃ for 4.5 h and the recombinant fusion protein was identified dy SDS PAGE and western blot.ResultsThe recombinant fusion protein band with relative molecule mass of 37 kD was shown on SDS PAGE gel. The band amounted to 25% of total bacteria protein. Western blot analysis showed that the band could react with anti IL 11 antibody. In addition,IL 11 could be obtained after TNFHis IL 11 was lysed with hydroxylamine.ConclusionTNFHis IL 11 fusion expression vecter was successfully constructed.