大鼠诱导型一氧化氮合酶短发夹环RNA质粒的构建及鉴定

目的：构建编码诱导型一氧化氮合酶（iNOS）Jfl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。

Construction and Identification of Plasmid Containing Short Hairpin RNA of Inducible Nitric Oxide Synthase of Rats

Objective：To construct a plasmid expression vector coding for the short hairpin RNA（shRNA） targeting iNOS promoter,for the use of gene activation in the treatment of erctile dysfunction in preparation. Methods：According to rat iNOS promoter sequence, were designed and synthesized shRNA oligonucleotides, annealed to form double-stranded and cloned into the vector Pdc316-EGFP-U6, recombinant plasmid, for PCR identification and sequencing. Results：PCR identification and sequencing showed that the recombinant plasmid was successfully con- structed. Conclusions：Tbe plasmid expression vectors coding for shRNA targeting iNOS promoter have been constructed successfuUy,established a favourable foundation for further explore gene therapy of erectile dysfunction.