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利用RT PCR法从小鼠肝组织扩增出Endostatin基因 ,重组入 pUC1 9质粒。经测序后 ,将片段重组入大肠杆菌表达载体 pDH并用温度诱导表达。将表达的蛋白经纯化及复性后直接注入大鼠C6胶质瘤模型内 ,检测其对胶质瘤生长的抑制作用。结果 ,在小鼠肝组织中成功扩增到Endostatin基因 ,测序与报道序列一致。重组进 pDH后经温度诱导表达 ,在SDS PAGE电泳上得到一条新的蛋白表达带 ,分子量为 2 0kD。纯化及复性后的蛋白能明显抑制小鼠体内胶质瘤的生长 ,为利用Endostatin进行脑胶质瘤基因治疗奠定了实验基础。
Endostatin gene was amplified from mouse liver tissue by RT PCR and recombined into pUC19 plasmid. After sequencing, the fragment was recombined into the E. coli expression vector pDH and induced by temperature. The expressed protein was purified and refolded and then directly injected into rat C6 glioma model to detect its inhibitory effect on glioma growth. As a result, Endostatin gene was successfully amplified in mouse liver tissue, and the sequence was consistent with the reported sequence. Recombinant pDH after induced by temperature, SDS PAGE electrophoresis to obtain a new protein expression zone, the molecular weight of 20kD. The purified and renatured protein can significantly inhibit the growth of glioma in mice and lay the experimental foundation for the gene therapy of glioma with Endostatin.