人端粒酶逆转录基因siRNA表达质粒的构建和鉴定
onstruction of shRNA expression vector system targeting hTERT gene plasmid
目的：构建含人端粒酶逆转录酶（Human telomerase reverse transcriptase，hTERT）基因的siRNA表达载体，为以端粒酶为靶点的基因治疗肺癌研究奠定基础。方法：化学合成能编码hTERT基因的短发夹RNA（shRNA）序列的寡核苷酸，退火处理克隆到pGenesil-1U6 shRNA表达载体相的U6RNA聚合酶Ⅲ（PolⅢ）启动子的下游，构建pGenesil-hTERT重组质粒。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pGenesil-hTERT载体，为进一步对其端粒酶活性的抑制作用打下基础。
Objective: To construct the expressing vector of small hairpin RNA (shRNA) targeting human telomerase reverse transcriptase (hTERT) gene for future study of Lung Cancer’s gene therapy. 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 plasmids pGenesil-hTERT. Results: It was demonstrated that shRNA template targeting hTERT gene had been inserted at the expected site and the insertion sequence was perfectly corrected. Conclusion: RNAi expression vector targeting hTERT gene has been constructed success fully and will be used as a useful method to develop specific hTERT- silencing therapeutics.
刘东雷、刘全、韩颖超、王建军、郑庆丰、严会志
肿瘤学生物科学研究方法、生物科学研究技术基础医学
RNA干扰短发夹RNA人端粒酶转录酶
RNA interferencesmall hairpin RNAhTERT
刘东雷,刘全,韩颖超,王建军,郑庆丰,严会志.人端粒酶逆转录基因siRNA表达质粒的构建和鉴定[EB/OL].(2010-02-05)[2025-05-22].http://www.paper.edu.cn/releasepaper/content/201002-308.点此复制
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