内皮抑素基因的获取及其分泌型质粒构建的研究

Study on Obtaining Endostatin Gene and the Construction of Secretion Plasmid Vector

目的 :通过分离、克隆、重组出内皮抑素基因并测定其基因序列 ,探讨单独或联合其它血管抑制因子基因及其表达产物经纯化作为生物制品可用于肿瘤治疗的研究。方法 :采用RT_PCR技术自肝癌组织中扩增出内皮抑素目的基因片段 ,采用pEZZ18为分泌型质粒载体构建含内皮抑素基因的重组质粒并在大肠杆菌DH5α内进行表达。结果 :成功获得内皮抑素基因并经序列测定证实。序列分析显示与已发表的内皮抑素基因无明显差异。克隆至 pEZZ18载体构建成功含内皮抑素基因的分泌型重组质粒。结论 :构建的重组质粒具有外分泌特性 ,为成功分泌表达内皮抑素、快速纯化及临床应用打下良好基础。

To explore the effects of lonely use or combined with other genes of angiogenetic inhibitors and endostatin in treating tumors by isolating,cloning,and sequencing endostatin gene.Methods:RT_PCR was used to amplify endostatin gene from hepatoma tissue,and pEZZ 18 was used to be secretable plasmid vector to construct the recombinant plasmid containing endostatin gene expressed in Ecoli.DH5α.Results:Endostatin gene was successfully obtained proved by sequencing.There was no significant difference in endostatin gene compared with the reported sequence.Conclusion:The constructed recombinant plasmid is secretable,and it is useful in expressing endostatin,purifying,and clinical use.