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

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 .