cblb502蛋白原核表达及辐射防护作用验证 - 中华放射医学与防护杂志.pdfVIP

·基础研究· CBLB502蛋白原核表达及辐射防护作用验证 王治东 葛常辉 许望翔 詹轶群 汤立宣 曹萌萌 杨晓明 【摘要】 目的原核表达并纯化TLR5受体的激动剂CBLB502蛋白，验证其辐射防护作用。 方法采用pET28b原核表达系统表达CBLBS02蛋白，利用Ni．NTA亲和层析柱亲和层析纯化 CBLB502蛋白，经报告基因检测方法结合动物实验评价纯化蛋白活性。结果成功表达并纯化 CBLBS02蛋白，纯度可达95％以上。能够激活NF．KB报告基因，可以提高9Gy照射小鼠存活率，对 照组小鼠全都死亡，CBLB502蛋白预防组全部存活。结论成功表达并纯化CBLB502蛋白。并验证 其对小鼠的辐射防护作用。 【关键词】TLR5；CBLB502蛋白；辐射防护 ofCBLBS02In andYermcationofradiation erect佴M|I＼rGZJIi． Expression prokaryoeytes protective 而增，GEChang-hui，XUWang-xiang，ZHANn·qun，TANGLi-算u口n，CAOMeng·meng，埘^留Xiao· InstituteRadiationMediation of 100850，China ming．Be聊ng Medicine，Be蚵ng Correspondingauthor：YANGXiao—ruing，Email：xiaomingyang@sina．corn Toobtainthe CBLB502asan ofT0ulike 5 【Abstract】0bJectivepurified proteinagonist receptor in andtovalidateitsradiation effect．Methods WaS prokaryocytes protective pET28bexpressionsystem usedto CBLB502 theNi．NTA and was columnusedto express protein．and agarosepurification purify CBLB502 radiation effectofCBLB502wasevaluatedNF．KB protein．Theproective by reportergeneassay and mice CBLB502WaS and couldactivateNF- experiments．Resultsproteinexpressedpurified。which xB invitroand the ofirradiatedmice 9 reportergene improvedlivability by Gy～．ray．Coneluslons CBLB502couldbe and has protein successfuHyexpressed system．It purifiedusin8prokaryoficexpression theexcellent effectinmice． radiation protective words】Toll-like5； CBLB502 R8diation 【Key receptor protein； protection Toll-like receptor(TLR)家族是一类参与病原微