Objective:To construct dopamine D1 receptor (DRD1) expression interference vectors to study the role of DRD1 in nerve cells and lay a foundation for drug development in anti-convulsion.Methods:Based on DRD1 gene sequence in GenBank,10 interfere vectors of DRD1 were designed.Liposomal was used to transfect NG-108-15 and the transfect effect was assayed by GFP.With realtime PCR and Western blot,the DRD 1 expression was detected.Results:The 10 constructed interfere vectors transfected into NG-108-15 cells by liposomal method and inhibited DRD1 mRNA and protein expression.DRD1 mRNA expression in NG-108-15 cells transfected with pGPU6-GFP-Neo-si-DRD1-5 was the lowest whereas DRD1 protein expression in NG-108-15 cells transfected with pGPU6-GFP-Neo-si-DRD 1-1,-2,-6,-7 was the lowest.Conclusion:DRD 1 expression interference vector is successfully constructed.%目的:构建多巴胺D1受体(DRD1)表达干扰载体,为研究DRD1在神经细胞中的作用及抗惊厥药物打下基础.方法:根据GenBank中DRD1基因序列,设计10个干扰序列并构建DRD1干扰载体.对NG-108-15进行脂质体转染后,GFP标记检测转染效果.Real-time PCR和Western印迹检测NG-108-15中DRD1的表达量.结果:构建的10个干扰载体均能采用脂质体法转染到NG-108-15细胞中.转染后NG-108-15中DRD1 mRNA和蛋白的表达均明显下降.其中转染pGPU6-GFP-Neo-si-DRD 1-5载体的NG-108-15中DRD1 mRNA表达水平最低,而转染pGPU6-GFP-Neo-si-DRD1-1,-2,-6,-7载体的NG-108-15中DRD1蛋白表达水平最低.结论:成功构建了DRD1表达干扰载体.
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