人白细胞介素18基因化学合成及其在大肠杆菌中高表达

目的设计并合成人白细胞介素18(hIL-18)基因,在大肠杆菌T7表达系统中高表达hIL 18。方法根据大肠杆菌的偏爱密码子和翻译起始区的二级结构,设计并人工合成了hIL-18的基因。将新合成基因克隆到载体pMFT7-5 ,经转化JM10 9(DE3) ,得到工程菌JTI。结果经IPTG诱导表达,SDS-PAGE结果表明目的蛋白质占总蛋白质的30 % ,具有免疫学活性。结论实现hIL基因在大肠杆菌中的高效表达

Chemical synthesis of human interleukin-18 gene and its high-level expression in E. Coli

PurposeTo realize the high-level expression of human interleukin-18 in T7 expression system in E.coli by designing and synthesizing human interleukin-18 gene.MethodsNew human interleukin gene was designed and synthesized,according to the preferred codons of E.coli and the secondary structure of translation initiation region. The expression vector,pMFT7-IL18,was constructed by cloning the gene into the vector pMFT7-5. The engineered strain,JTI,constructed by transforming pMFT7-IL18 into JM109(DE3),was used to express target protein.Results30% expression ratio was achieved after induction by IPTG,according to the consequence of SDS-PAGE analysis. Immunology activity was identified by western blotting.ConclusionHigh-level expression of human interleukin-18 in E.coli was achieved.