论文部分内容阅读
目的构建胃癌新生血管导向性肽GX1与新型人肿瘤坏死因子(GX1-rmhTNF)基因的原核表达载体,表达GX1-rmhTNF蛋白。方法利用基因工程方法将胃癌血管特异性结合肽融合在新型人肿瘤坏死因子α的氨基末端,构建GX1-rmhTNF原核表达载体,在大肠杆菌进行表达。对表达产物进行SDS-PAGE电泳分析和Westernblot检测分析。结果成功构建了GX1-rmhTNF重组融合表达质粒,表达的蛋白经SDS-PAGE电泳分析,在相对分子质量(Mr)约18000处出现了1条新生的蛋白条带,该表达蛋白具有与TNFα单克隆抗体特异性的结合能力。结论成功构建了GX1-rmhT-NF基因的原核表达载体并在原核细胞中表达了该产物,为下一步GX1-rmhTNF的纯化奠定了基础。
Objective To construct prokaryotic expression vector of GX1 and novel human tumor necrosis factor (GX1-rmhTNF) gene and to express GX1-rmhTNF protein. Methods The fusion protein of vascular specific binding peptide of gastric cancer was fused to the amino terminal of novel human tumor necrosis factor α by genetic engineering method. The prokaryotic expression vector GX1-rmhTNF was constructed and expressed in E. coli. The expression products were analyzed by SDS-PAGE electrophoresis and Western blot analysis. Results Recombinant GX1-rmhTNF fusion protein was successfully constructed. The expressed protein was analyzed by SDS-PAGE. One new protein band appeared at about 18000 molecular weight (Mr) Antibody-specific binding ability. Conclusion The prokaryotic expression vector of GX1-rmhT-NF gene was successfully constructed and expressed in prokaryotic cells, which laid the foundation for the further purification of GX1-rmhTNF.