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Construction and expression of mouse endostatin in eukaryotic expression vector
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 9
- DOI:
- 10.3321/j.issn:1000-2790.2000.09.005
- Key Word:
- endostatin;真核表达;胶质瘤
Abstract： 目的构建小鼠内皮抑素(endostatin) cDNA的真核表达载体并将其在体外培养的脑胶质瘤细胞内表达.方法将带有分泌信号的小鼠endostatin cDNA克隆入真核表达载体pcDNA3,构建CMV启动子控制的载体pcDNA-sEndo,并将上述载体转染体外培养的脑胶质瘤细胞(SHG44),采用Western Blot鉴定其表达,应用牛微血管内皮细胞抑制实验检测其活性.结果成功地构建了真核表达载体pcDNA-sEndo并转染入脑胶质瘤细胞,获得的分泌型endostatin可以明显抑制牛毛细血管内皮细胞增殖(ED50=200 μg.L-1).结论获得有表达功能活性的endostatin真核表达载体,为脑胶质瘤的抗血管生成基因治疗提供了必要的条件.
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