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人醛缩酶A干扰RNA真核表达载体的构建

Construction of Eukaryotic Expression Vectors of Human Total ALDOA
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摘要 目的 构建4个人醛缩酶A （aldolaseA,ALDOA）公共序列的干扰RNA真核表达载体,转染人脑神经胶质细胞U251细胞,为醛缩酶A成为人脑肿瘤标志物提供实验基础和有价值的资料.方法 ①构建4个新靶点醛缩酶A干扰RNA真核表达载体并用双酶切和测序鉴定;②用脂质体法瞬时转染上述4个载体到U251细胞株.结果 ①成功构建了4个新靶点的醛缩酶A干扰RNA真核表达载体即pGPU6/GFP/Neo-ALDOA-1,pGPU6/GFP/Neo-ALDOA-2,pGPU6/GFP/Neo-ALDOA-3和pGPU6/GFP/Neo-ALDOA-4;②在U251细胞中有绿色荧光蛋白表达.结论 成功构建并筛选出效果最好的新靶点MBP干扰RNA真核表达载体. Objective Four new eukaryotic expression vectors of RNA interference specific for ALDOA that were transfectedto inhibit the expression of ALDOA in U251 cell were constructed via the technique of RNA interference. Research the roleof MBP and provide valuable theoretical basis for neonatal brain injury. Methods （1）Four new eukaryotic expression vectorsof RNA interference specific for ALDOA were constructed, whose names were pGPU6/GFP/Neo-ALDOA-1, pGPU6/GFP/Neo-ALDOA 2, pGPU6/GFP/Neo ALDOA-3 and pGPU6/GFP/Neo-ALDOA-4. （2）U251 cell line was transfected with fournew vectors above via oligofectamine. Results （3）New targets of eukaryotic expression vector of RNA interference specificfor ALDOA was constructed. OGreen fluorescent protein expressed in U251 cells. Conclusion Human U251 cell line thatwas transfectcd by new Eukaryotic expression vectors of ALDOA siRNA.
作者 董莉 呼格吉乐 易青 萨日娜 王悦喜
机构地区 内蒙古医科大学附属医院检验科
出处 《现代检验医学杂志》 CAS 2014年第2期38-40,43,共4页 Journal of Modern Laboratory Medicine
基金 内蒙古自治区教育厅高等学校科学研究项目重大课题（NJZZ13411）.
关键词 干扰RNA 醛缩酶A 神经胶质细胞 U251 interference RNA ALDOA neuroglial call U251
分类号 Q786 [生物学—分子生物学]
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参考文献12

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现代检验医学杂志
2014年 第2期

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