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目的:构建编码诱导型一氧化氮合酶(iNOS)启动子的shRNA质粒表达载体,为利用基因激活技术治疗勃起功能障碍的研究做准备。方法:根据大鼠iNOS启动子序列设计并合成shRNA寡核苷酸片段,退火形成双链并克隆进入载体pDC316-EGFP-U6,构建重组质粒,并进行PCR鉴定以及测序分析。结果:PCR鉴定以及测序证实重组质粒构建成功。结论:成功构建了靶向iNOS基因的shRNA质粒表达载体,为进一步探索勃起功能障碍基因治疗奠定了基础。
OBJECTIVE: To construct shRNA plasmid expression vector encoding inducible nitric oxide synthase (iNOS) promoter and to prepare gene therapy for erectile dysfunction. Methods: According to the rat iNOS promoter sequence, an oligonucleotide fragment of shRNA was designed and synthesized. The double stranded DNA was annealed and cloned into the vector pDC316-EGFP-U6 to construct a recombinant plasmid. The recombinant plasmid was identified by PCR and sequenced. Results: PCR identification and sequencing confirmed that the recombinant plasmid was successfully constructed. Conclusion: shRNA plasmid expression vector targeting iNOS gene was successfully constructed, which laid the foundation for further exploration of gene therapy of erectile dysfunction.