抗肿瘤的人重组内皮抑素基因的克隆及表达

Ooning and Expression of Antitumor of Recombinant Human Endostatin Gene

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 and express recombinant human endostatin, (HES). Methods The lies gene was isolated with RT- PCR from human placenta tissue total RNA. Then, the 570 bp endostatin gene fragment was cloned into pUC18 vector for sequencing. It was expressed and purified by Xpress system. Results The DNA sequence of cloned hes gene was similar to that reported previously, and fusion protein of recombinant hes expression was above 40% of total bacterial proteins. Conclusions The hes gene and its recombinamt with higt level expression were obtained. Success of cloning and expression of human endostatin gene will provide tool for high level expression of it and further clinical application as antitumor drug.