肿瘤坏死因子受体相关因子家族成员间相互作用的初步研究  

Prime study of the protein-protein interaction among tumor necrosis factor receptor-associated factors

作　　者：周扬帆[1] 王晓辉[2] 唐刘君[2] 李长燕[2] 王敏[1] 杨晓扬[2] 杨晓明[2] 汪思应[1] 

机构地区：[1]安徽医科大学病理生理教研室,合肥230032 [2]军事医学科学院放射与辐射医学研究所,北京蛋白质组研究中心,北京102206

出　　处：《安徽医科大学学报》2011年第10期989-993,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家重大科学研究计划(973计划)资助项目(编号:2011CB910600)

摘　　要：目的探讨肿瘤坏死因子受体相关因子(TRAFs)各成员间的相互作用及各成员间的关系。方法利用PCR技术扩增出TRAFs;构建到pDBLeu、pPC86载体上,分别形成诱饵、猎物质粒;转化,扩增酵母菌;将诱饵、猎物菌液结合酵母双杂交阵列组合(Mating)技术筛选相互作用。最后用免疫共沉淀技术进行验证。结果成功构建TRAFs的酵母双杂交载体,酵母双杂交Mating技术筛选出11对相互作用,免疫共沉淀技术验证出,TRAF1和TRAF2、TRAF6有相互作用。结论 TRAFs各成员间存在着相互联系;TRAF1通过和TRAF6的相互作用可能直接影响TRAF6介导的通路。Objective To identify the protein-protein interaction among tumor necrosis factor receptor-associated factors（TRAFs） and reveal their relation.Methods Full-length cDNA of TRAFs were obtained by polymerase chains reaction（PCR）,then cloned into pDBLeu vector and pPC86 vector,named bait vector and prey vector which were transformed into different yeast cells.Then the baits combined with the preys freely.The positive results were verified by co-immunoprecipitation.Results The relevant vectors of TRAFs were successfully cloned.11 interactions were obtained by Y2H screening.TRAF1 was confirmed to interact with TRAF2,TRAF6 by co-immunoprecipitation assay.Conclusion TRAFs members exist linkage.TRAF1 may effect the signaling pathway which is induced by TRAF6.

关 键 词：酵母双杂交Mating TRAFs 相互作用 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R341.32[医药卫生—基础医学]