人白细胞介素-6成熟肽基因重组及其在大肠杆菌中的表达

Recombination of IL-6 gene and Expression of it in E. coli

将PCR扩增的人白细胞介素-6（hIL-6）成熟肽基因与PJLA502表达载体重组，在大肠杆菌中高效表达出rhIL－6。经SDS－PAGE分析，rhIL－6表达量占菌体可溶性蛋白的40％以上。经免疫印迹证实，rhIL－6具有良好的抗原特异性。复性后，用MTT法测定IL－6依赖株7TD1细胞增殖活性，rhIL－6比活性达2．5×10~6u／mg蛋白。

The gene which seems to encode the mature human IL-6 was amplified by PCR and inserted into expression vector pJLA502. Hybridization in situ and restriction endonuclea.e analysis showed that recombinant clone contained the coding region for mature hIL-6. The mature hIL-6 was expressed in E. coli and the molecular weight of expressed protein was approximately 21 KD by SDS-PAGE. More than 40% of somatic soluble protein of rhIL-6 was expressed. Western blot revealed that 21KD protein showed specific antigenicity. The specific activity of rhIL-6 measured by MTT method could reach to 2.5 × 10~6 u/mg protein.