重组hIL-24原核表达载体的构建及表达

采用基因克隆的方法构建人白细胞介素(hIL-24)基因的原核表达载体pET-28a(+)-hIL24,在大肠杆菌中诱导表达重组蛋白rhIL-24,SDS-PAGE和Western blotting分析蛋白表达,探讨其最佳诱导表达条件.结果表明:hIL-24基因的表达产物分子质量约25 ku,且以包涵体的形式存在于超声裂解后的沉淀中,重组蛋白rhIL-24可特异性被鼠抗人IL-24单克隆抗体识别.当IPTG终浓度为0.5 mmol·L~(-1)、37℃诱导8 h时,其表达量最高.hIL-24基因在该原核表达系统中的成功表达,为其生物学功能的研究奠定了基础.

Construction and expression of the recombinant prokaryotic expression vector with hIL-24

In this study,a recombinant expression vector of human interleukin-24(hIL-24)gene,named pET-28a(+)-hIL24,was constructed by gene cloning.The recombinant protein hIL-24 was expressed in E. coil and analyzed by SDS-PAGE and Western blotting.The proper inducing conditions of its expression were explored.The results showed that the recombinant protein was about 25 ku and mainly existed in the precipitation after sonication.Western blotting proved that the recombinant protein IL-24 was recognized by mouse anti-human IL-24.SDS-PAGE analysis indicated that the best expression condition was 0.5 mmol·L~(-1) IPTG for 8 h at 37℃.The successful expression of hIL-24 in the prokaryotic expression system is helpful for the further study of its biological function.