GLT-1基因siRNA真核表达载体的构建及其在大鼠神经胶质细胞中的抑制效应

发布时间：2018-03-07 22:11

  本文选题：GLT-基因　切入点：RNA干扰　出处：《生物技术通讯》2007年01期 　论文类型：期刊论文

【摘要】：目的:研究特异性siRNA对大鼠神经胶质细胞中GLT-1基因的阻抑效果。方法:根据GLT-1基因的序列特点和RNAi设计原则,设计其shRNA的核苷酸片段。退火后将其克隆入pSupressorNeo,构建可表达大鼠GLT-1基因siRNA的重组真核表达质粒pSuppressorNeo-GLT-1;利用脂质体法将其转染神经胶质细胞后,用RT-PCR、Western印迹及免疫荧光法等方法检测转染的神经胶质细胞中GLT-1基因的表达水平。结果:瞬时转染的神经胶质细胞中GLT-1基因的表达受到明显抑制,GLT-1蛋白含量明显下降。结论:pSuppressor-Neo-GLT-1质粒构建成功,瞬时转染神经胶质细胞后可以明显抑制GLT-1基因的表达。
[Abstract]:Objective: to study the inhibitory effect of specific siRNA on GLT-1 gene in rat glial cells. Methods: according to the sequence characteristics of GLT-1 gene and the principle of RNAi design, The nucleotide fragment of shRNA was designed and cloned into pSupresor Neo. after annealing, the recombinant eukaryotic expression plasmid pSuppressorNeo-GLT-1 was constructed and transfected into glial cells by liposome method, and the recombinant eukaryotic expression plasmid pSuppressorNeo-GLT-1 was constructed. The expression level of GLT-1 gene in transfected glial cells was detected by RT-PCR Western blot and immunofluorescence. Results: the expression of GLT-1 gene in transient transfected glial cells was significantly inhibited and the content of GLT-1 protein was significantly decreased. Conclusion the plasmid pSuppressor-Neo-GLT-1 was constructed successfully. Transient transfection of glial cells significantly inhibited the expression of GLT-1 gene.
【作者单位】： 第四军医大学唐都医院放射科 解放军第451医院放射科
【分类号】：Q78

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