人TRAIL胞外区原核可溶性表达及活性鉴定  

Expression the tracellular domain of human TRAIL and identification of its activity

作　　者：鲍应环 程相树[1] 刘峰涛[1] 

机构地区：[1]河南大学医学院细胞与分子免疫学实验室,河南开封475004

出　　处：《河南大学学报（医学版）》2014年第2期117-119,共3页Journal of Henan University:Medical Science

基　　金：河南省医学科技攻关计划项目资助(WKJ2007-021)

摘　　要：目的获得具有生物活性的肿瘤坏死因子相关凋亡诱导配体(TRAIL)胞外段蛋白。方法根据大肠杆菌密码子偏爱性要求,设计合成编码TRAIL胞外段的DNA序列,构建成pET30a-TRAIL胞内融合表达质粒,重组质粒转化表达宿主E.coli BL21。在不同温度、不同浓度的IPTG条件下诱导表达TRAIL,SDS-PAGE分析表达产物,MTT法检测产物活性。结果构建的工程菌株表达29KD的可溶性TRAIL融合蛋白,能诱导Jurkat细胞凋亡。结论成功表达了人TRAIL胞外段蛋白,为进一步研究提供基础。Objective To obtain the extracellular domain of tumor necrosis factor-related apoptosis-inducing ligand （ TRAIL） with activity . Methods We designed and synthesized DNA encoding the TRAIL extracellular DNA sequences according to the coding preference of the E .coli and constructed the prokaryotic expression vector ：the pET30a TRAIL fusion expression plasmid . The recombinant plasmid was transformed into the expression host E .coli BL21 to induce expression under different temperature conditions , different concentrations of IPTG and alcohol . After inducing expression we used the SDS-PAGE method to analysis of expression product and used the MTT method to assay the activity of expression product .Results The constructed engineering bacteria expressed a 29KD soluble TRAIL fusion protein , and this protein can induce Jurkat cell apoptosis .Conclusion We successful express the extracellular domain of human TRAIL protein with activities .

关 键 词：原核表达 JURKAT细胞 凋亡 

分 类 号：Q782[生物学—分子生物学]