STAT3基因 siRNA 表达载体的构建及鉴定

摘要

Objective This study used RNA interference technology,selected STAT3 gene as the tar-get,and then design and construction a recombinant vector and then identification the gene sequence analy-sis.Methods Design a DNA sequence with a small hairpin structure,PCR amplification a siRNA expression cassettes,cloning vector to construct recombinants in psiLentGeneTM into E.coli DH5αstrain,extract plas-mid after enzyme digestion sequencing.Results Recombinant plasmid by EcoR Ⅴ enzyme,electrophoresis and Sequencing analysis showed that the inserted sequence is correct,the recombinant plasmids were success-fully constructed.Conclusion The successful construction of STAT3 targeted RNA interference recombi-nant,provides some new methods for gene therapy of cancer research at the molecular level,cellular level and the overall level of.%目的：利用RNA干扰技术，以信号转导和转录激活因子3（STAT3）为靶基因，设计构建重组体，并进行序列分析鉴定。方法设计一条有小发夹结构的 DNA 序列，聚合酶链反应（ PCR）扩增合成 siRNA 表达框架（SECs）表达框架，克隆至载体 psiLentGeneTM 中构建重组体，转化 E.coli DH5α菌株，提取质粒进行酶切鉴定后测序分析。结果重组质粒经 EcoRⅤ酶切、电泳、测序分析表明插入序列正确，重组质粒构建成功。结论成功构建了 STAT3靶向 RNA 干扰重组体，为肿瘤的基因治疗在分子水平、细胞水平和整体水平的研究提供一些新方法。