【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。更多还原

【Abstract】 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 pET30aTRAIL fusion expression plasmid.The recombinant plasmid was transformed into the expression host E.coli BL21to 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.更多还原