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目的 克隆抗肿瘤因子内皮抑素基因并表达内皮抑素蛋白重组人内皮抑素(human endosbtatin,HES)。方法 从人胎盘组织中分离总RNA,用RT-PCR法扩增出570bp的DNA片段,经序列测定证实为内皮抑素基因,并成功地克隆到pUC18质粒载体中,用Xpress系统体外表达出内皮抑素蛋白并纯化成功。结果 克隆的HEScDNA其序列和国外报道一致;构建了重组HES融合蛋白表达菌株,经SDS-PAGE等分析,表达目的蛋白达细菌总蛋白的40％以上。结论 通过HES基因克隆和表达的研究与探讨,得到了HES的基因克隆和高效表达菌株,为内皮抑素在临床治疗恶性肿瘤奠定了基础。
Objective To clone the anti - oncogene endostatin gene and express the endostatin protein recombinant human endosbtatin (HES). METHODS: Total RNA was isolated from human placenta tissue. The 570 bp DNA fragment was amplified by RT-PCR. The endostatin gene was confirmed by sequencing and successfully cloned into pUC18 plasmid vector. Endothelial Stroma protein is successfully purified. Results The cloned HES cDNA sequence was consistent with that reported in foreign countries. The recombinant HES fusion protein was constructed and analyzed by SDS-PAGE. The expressed protein reached 40% of total bacterial protein. Conclusions The cloning and expression of HES gene were studied. The cloned and highly expressed HES strains were obtained, which laid the foundation for the clinical treatment of malignant tumors by endostatin.