人白介素24(IL-24)的表达与纯化及其初步活性评价  

Expression,Purification and Preliminary Activity Evaluation of Interleukin 24 in Escherichia coli

作　　者：于明磊[1] 依莫安[1] 张家骊[1] 陈蕴[1] 金坚[1] 

机构地区：[1]江南大学药学院,江苏无锡214122

出　　处：《药物生物技术》2013年第4期314-317,共4页Pharmaceutical Biotechnology

摘　　要：原核表达IL-24蛋白并获取高纯度目的蛋白。选择大肠杆菌作为宿主菌,将已构建的重组表达质粒pET21a(+)/IL-24转化入大肠杆菌BL21(DE3)感受态细胞,获得重组表达IL-24的工程菌。通过对工程菌进行5 L发酵罐高密度发酵培养,获取目的蛋白,表达产物经SDS-PAGE和Western blot鉴定。目的蛋白通过阴、阳离子交换层析联用进行纯化。高密度发酵后单位体积目的蛋白的表达量高达1 800 mg/L。经阴、阳离子交换层析联用纯化后的回收率达20%,纯度达到95%。获得高纯度IL-24,为进一步研究其生物学活性及临床价值奠定了基础。The aim of the study is to express and purify interleukin-24.According to the expression features of E.coli,a recombinant expression plasmid pET21a（＋） / IL-24 was transformed into E.coli BL21（DE3）.The expression was carried out at 5L fermenter with harvest cell weight of 186.32gm（Max level achieved 255.5gm）.In high cell density fermenter this expression level was raised to 1800mg / L.Recombinant human interleukin 24 expression was carried out without any tag.The insoluble inclusion body obtained from wet cell paste was further processed for purification.Protein purification designed in two steps,Q-sepharose and SP-sepharose fast flow columns and the recovery of specific protein was 20 %,and the purify was up to 95%.The expressed proteins were quantified using SDS-PAGE,and the molecular weight of the protein was about 18.5 k which is consistent with the theoretical molecular weight.A lot of IL-24 was obtained.This laid a foundation for further studies on biological activity and clinical value of interleukin-24.

关 键 词：IL-24 发酵 纯化 复性 

分 类 号：Q51[生物学—生物化学]