摘要 :目的 构建端粒酶逆转录酶(hTERT)基因的RNA干扰(RNAi)表达载体。 方法 化学合成能编码针对hTERT基因的短发夹RNA(shRNA)序列的寡核苷酸，退火处理后克隆到pSilencer1.0-U6shRNA表达载体的U6RNA聚合酶Ⅲ(PolⅢ)启动子的下游，构建重组RNAi质粒pSliencer-hTERT。 结果 经酶切电泳及测序分析证实，目的序列成功插入到预计位点。 结论 已成功构建pSliencer-hTERT载体，为进一步研究其对端粒酶活性的抑制作用打下基础。

    关键词 :RNA干扰;短发夹RNA;人端粒酶转录酶;重组质粒;

    Construction of shRNA expression vector system targeting hTERT gene plasmid

    MAO Li-jun，ZHENG Jun-nian，LI Wang，et al

    (Department of Urology，Affiliated Hospital of Xuzhou Medical College，Xuzhou，Jiangsu221002，China)

    Abstract:Objective To construct the expressing vector of small hairpin RNA(shRNA)targeting human telomerase re-verse transcriptase(hTERT)gene.Methods Two template DNA oligonucleotides for shRNA expression targeted hTERT gene were chemically synthesized.The annealed shRNA templates were processed to produce and identify the recombinant RNAi plas-mid sPilencer-hTERT.Results It was demonstrated that shRNA template targeting hTERT gene had been inserted at the ex-pected site and the insertion sequence was perfectly correct.Conclusion RNAi expression vector targeting hTERT gene has been constructed successfully and will be used as a useful method to develop specific hTERT-silencing therapeutics. ......