[摘 要] 目的: 应用人U6启动子构建RNA干扰载体pUC19NU，具有新霉素抗性且能够在真核细胞中复制，成为基因功能和基因治疗研究等领域的RNAi技术的工具。方法: 通过PCR从人类基因组中得到人U6 snRNA启动子，并针对增强绿色荧光蛋白(EGFP)和p53蛋白的基因，分别设计了两段可以转录出短发夹环状RNA的DNA模板序列，连入pUC19中，构建出RNA干扰质粒pUC19NU，分别得到质粒载体pUC19NUE 和 pUC19NUP。 结果: 以EGFP 和p53为靶基因的干扰实验证明，两种质粒载体分别转染HeLa细胞和KMB-17细胞，两种蛋白的表达皆受到有效抑制。结 论: 该质粒能够有效地干扰相应内源性和外源性靶基因的表达，可以用于RNAi技术的研究。

    [关键词]RNA干扰; U6启动子; 绿色增强荧光蛋白; p53; 质粒

    Construction and detection of the plasmid vector with the human U6 snRNA promoter for RNA Interference

    XIA Chun-xiang， XIE Man-xia， CHU Li-hui

    (Department of Clinical Laboratories，the Fourth People′s Hospital of Huai'an， Huai'an Jiangsu 223300， China) [Abstract] Objective: To construct a vector to get the siRNAs for RNAi. Methods: The human U6 snRNA promoter from human genome was amplified by PCR ，cloned into the reconstructed plasmid to gen- erate the vector for RNAi ，which called pUC19NU. To test the RNAi responses directed by the vector， de- signed two DNA templates that can be transcripted into shRNA against enhanced green fluorescent protein (EGFP) or p53 protein in cells and inserted them into pUC19NU respectively ......