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Construction and identification of antisense VEGF165 eukaryotic expression vector
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 10
- DOI:
- 10.3321/j.issn:1000-2790.2000.10.002
- Key Word:
- 血管内皮细胞生长因子;真核表达载体;构建;胶质瘤,表达
Abstract： 目的进行抗血管生成治疗人脑胶质瘤实验,构建和鉴定携带人反义VEGF165基因的真核表达载体. 方法将VEGF165 cDNA反向克隆入pcDNA3,构建CMV启动子控制的真核表达载体pcDNA-AVEGF165,用酶切鉴定结果;应用该载体转染人脑胶质瘤细胞SHG44,G418筛选阳性克隆;Western blot和免疫组化检测转染前、后瘤细胞的VEGF表达. 检测转染前、后瘤细胞的生物学性状. 结果获得反向构建的pcDNA-AVEGF165真核表达载体,VEGF165反义RNA阻断了SHG44细胞的VEGF表达,该细胞生物学性状不受外源基因的表达影响. 结论成功构建的反义VEGF165真核表达载体,为人脑胶质瘤的反义基因治疗提供了必要的条件.
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