重组CC家族趋化因子RANTES的原核表达及趋化活性分析  

Expression in E.coli and Chemotactic Activity Analyzing of Recombinant CC-chemokine RANTES

作　　者：孙荣华[1,2] 王应[1] 钟英诚[1] 马大龙[1] 

机构地区：[1]北京大学基础医学院分子免疫室 [2]Fax:[phone]

出　　处：《中国医学科学院学报》2001年第2期123-126,共4页Acta Academiae Medicinae Sinicae

摘　　要：目的 深入研究趋化因子 RANTES的结构与功能，为其进一步的应用研究打下基础。方法 利用 PCR技术扩增 RANTES成熟蛋白的编码区序列，将其克隆到原核表达载体，经表达纯化得到重组蛋白质纯品，利用趋化小室法测趋化活性。结果 成功扩增 RANTES cDNA编码区序列，在大肠杆菌中成功表达重组 RANTES蛋白，经纯化蛋白纯度达 95％以上，对外周血淋巴细胞（ PBL）， U937淋巴瘤细胞系，和 HEK293-CCR4稳定株有趋化活性，对 Jurkat细胞无明显趋化作用。结论 原核表达的重组 RANTES对多种细胞具有趋化活性，并建立了用培养细胞系检测趋化活性的模型，为下一步 RANTES的结构与功能研究打下了基础。Objective To further understand the structure and function of RANTES and provide basis for research for application. Methods PCR product corresponding to region encoding mature RANTES protein was cloned into E.coli expression vector; purified recombinant protein was obtained by heparin affinity chromatography and its chemotactic activity was determined by Boyden chamber. Results The recombinant RANTES was expressed in E.coli with high efficiency. Purified protein showed chemotactic activity to peripheral blood lymphocytes, U937 cells, and CCR4 stable-transfected HEK293 cells. Conclusion The purified recombinant protein exerted chemotactic activity on PBL and cultured cell lines, and we have established a experimental system for further study of structure and function of RANTES.

关 键 词：趋化因子 RANTES 原核表达 趋化活性 重组CC家族趋化因子 

分 类 号：R392.11[医药卫生—免疫学]