| 重组人IL-17A和IL-17F的制备方法及活性测定 |
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| 引用本文： | 贾军会,李慎涛,刘振龙,赵文明.重组人IL-17A和IL-17F的制备方法及活性测定[J].生物技术通报,2011(4). |
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| 作者姓名： | 贾军会 李慎涛 刘振龙 赵文明 |
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| 作者单位： | 首都医科大学免疫学系,北京,100069 |
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| 摘 要： | 人IL-17A和IL-17F具有很高的同源性,在炎症性疾病、自身免疫性疾病和肿瘤中都发挥着重要的作用,是当前研究的热点.应用原核表达系统在大肠杆菌BL21(DE3)中高效表达了人IL-17A和IL-17F;经培养条件的优化,未发现可溶性目的蛋白的表达,免疫印记分析显示,重组蛋白位于包涵体中;对包涵体进行洗涤、凝胶过滤层析纯化和柱上复性,获得重折叠的可溶性蛋白;随后用SDS-PAGE对蛋白样品进行了纯度分析、采用免疫印记和质谱的方法鉴定蛋白产物成分、用ME3T3-E1和RAW264.7两个细胞株对IL-17A、IL-17F的生物学活性进行测定.结果显示,柱上复性的方法制备的谊重组蛋白具有较高的纯度和活性.建立的重组人IL-17A和IL-17F的制备方法可为相关研究中细胞因子的大量应用提供参考.
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| 关 键 词： | 蛋白纯化 柱上复性 活性测定 |
Preparation of Recombinant Human IL-17A and IL-17F and Determination of Their Bioactivities |
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| Jia Junhui,Li Shentao,Liu Zhenlong,Zhao Wenming.Preparation of Recombinant Human IL-17A and IL-17F and Determination of Their Bioactivities[J].Biotechnology Bulletin,2011(4). |
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| Authors: | Jia Junhui Li Shentao Liu Zhenlong Zhao Wenming |
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| Institution: | Jia Junhui Li Shentao Liu Zhenlong Zhao Wenming(Immunology Department,Capital Medical University,Beijing 100069) |
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| Abstract: | Human IL-17A and IL-17F,two most important cytokines in inflame diseases,autoimmune diseases and cancer,have been studied extensively.For further research of their roles in Rheumatoid Arthritis(RA),IL-17A and IL-17F were expressed in E.coli and purification protocol was established.Both the IL-17A and IL-17F were expressed in E.coli BL21(DE3)in high yield in the form of inclusion bodies.The inclusion bodies were washed in PBS,TritonX-100,4 mol/L Urea subsequently,and then dissolved in lysis buffer.The denat... |
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| Keywords: | IL-17A IL-17F |
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