| 引用本文： |
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刘娜,严海东,李雪竹.MCP-1siRNA真核表达载体的构建及其对HKCMCP-1基因沉默效应的鉴定[J].同济大学学报（医学版）,2007,(2):42-46. [点击复制]
- .Construction of eukaryotic expression vector of siRNA specific for monocyte chemotacite protein-1 and charcterization of its efficiency in HKC[J].Journal of Tongji University(Medical Science),2007,(2):42-46. [点击复制]
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| 摘要: |
| 目的构建单核细胞趋化蛋白1(MCP-1)特异性小分子干扰RNA(siRNA)的真核表达载体,研究siRNA对人肾小管上皮细胞(HKC)MCP-1基因的沉默作用。方法采用基因克隆技术,将合成的MCP-1特异性siRNA插入真核表达载体pRNAT-U6/Neo中,构建MCP-1 siRNA真核表达载体。将pRNAT-MCP-1质粒以脂质体法转染HKC,24 h后应用实时定量RT-PCR技术检测HKC内MCP-1 mRNA水平的表达情况。结果成功构建MCP-1siRNA真核表达载体,转染MCP-1 siRNA的HKC,24 h后HKC内MCP-1 mRNA水平表达明显降低,抑制效率达90%以上。结论构建的RNA干扰真核表达载体能够高效抑制HKC的MCP-1基因表达,为肾间质纤维化的防治研究奠定了实验基础。 |
| 关键词: 单核细胞趋化蛋白1,小分子干扰RNA,真核表达载体,肾间质纤维化,人肾小管上皮细胞 |
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| 修订日期：2006-12-12 |
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| Construction of eukaryotic expression vector of siRNA specific for monocyte chemotacite protein-1 and charcterization of its efficiency in HKC |
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| Abstract: |
| Objective To construct the small interfering RNA(siRNA) eukaryotic expression vector specific for monocyte chemotacite protein-1(MCP-1) and to observe its silencing effection on MCP-1.Methods The expression vector of pRNAT-U6/Neo/MCP-1 siRNA was construct |
| Key words: monocyte chemotacite protein-1,small interfering RNA,eukaryotic expression vector,renal interstitial fibrosis,human renal tubular epithelial cells |
