小鼠白介素17受体A胞外段的表达、纯化及鉴定  

Expression,purification and identification of mIL-17RA_(aa32-322)

作　　者：魏衍财[1] 朱晨露[1] 陈艳红[1] 秦伟[1] 田洁[1] 袁靖[1] 陈娟[1] 吴晶晶[1] 汤新逸[1] Samuel Essien Baidoo 许化溪[1] 王胜军[1] 

机构地区：[1]江苏大学检验医学研究所,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2012年第4期282-285,共4页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(31170849;81072453;30972748);江苏省自然科学基金资助项目(BK2011472);江苏省普通高校研究生科研创新计划项目(CXLX11_0608);江苏省卫生厅医学科研基金资助项目(200952)

摘　　要：目的:分别克隆小鼠白介素17受体A胞外段(mIL-17RAaa32-322)及小鼠IgG2AFc(mIgG2AFc)基因,构建mIL-17RAaa32-322-mIgG2AFc融合蛋白原核表达载体,并在体外表达、纯化及鉴定。方法:利用基因重组技术构建pET32a(+)-mIL-17RAaa32-322-mIgG2AFc原核表达载体,并在E.coli Rosetta中表达mIL-17RAaa32-322-mIgG2AFc融合蛋白,经镍离子螯合层析分离纯化后,用SDS-PAGE电泳和蛋白质印迹法进行鉴定。结果:成功构建pET32a(+)-mIL-17RAaa32-322-mIgG2AFc原核表达载体,获得高效表达的融合蛋白,且蛋白质印迹证实为目的蛋白。结论:获得mIL-17RAaa32-322-mIgG2AFc融合蛋白,为进一步研究其生物学功能奠定基础。Objective： To clone the extracellular region of mouse interleukin-17 receptor（mIL-17RAaa32-322） and mouse IgG2AFc（mIgG2AFc）,construct the recombinant prokaryotic expression vector of mIL-17RA aa32-322-mIgG2AFc,and then to express,purify and identify the fusion protein.Methods： The prokaryotic expression vector pET32a（＋）-mIL-17RAaa32-322-mIgG2AFc was constructed by using recombinant DNA technology and expressed in E.coli Rosetta.Then mIL-17RAaa32-322-mIgG2AFc fusion protein was purified by nickel-chelating chromatography and identified by SDS-PAGE electrophoresis and Western blot assay.Results： The prokaryotic expression vector pET32a（＋）-mIL-17RAaa32-322-mIgG2AFc was successfully constructed,which can express fusion protein of high purity.The mIL-17RAaa32-322-mIgG2AFc was confirmed by Western blot.Conclusion： Recombinant prokaryotic expression vector pET32a（＋）-mIL-17RAaa32-322-mIgG2AFc was successfully constructed and mIL-17RAaa32-322-mIgG2AFc fusion protein was purified for the research and application of foundation in the treatment of autoimmune disease.

关 键 词：小鼠白介素17受体A 原核表达 纯化 鉴定 

分 类 号：R392.11[医药卫生—免疫学]