CAJ | 학술논문

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（T RA IL ）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29 KD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。
목적：획득구유생물활성적종류배사인자상관조망유도배체（T RA IL ）포외단단백。방법근거대장간균밀마자편애성요구，설계합성편마TRAIL 포외단적DNA서렬，구건성pET30a-TRAIL포내융합표체질립，중조질립전화표체숙주 E ．coli BL21。재불동온도、불동농도적IPTG조건하유도표체 TRAIL ，SDS-PAGE분석표체산물，MTT법검측산물활성。결과구건적공정균주표체29 KD적가용성TRAIL융합단백，능유도Jurkat세포조망。결론성공표체료인TRAIL포외단단백，위진일보연구제공기출。
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 .