STAT3基因siRNA表达载体的构建及鉴定

The Building and Identification of siRNA STAT3 Gene Expression Vector

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psi Lent GeneTM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。

Objective This study used RNA interference technology,selected STAT3 gene as the target,and then design and construction a recombinant vector and then identification the gene sequence analysis. Methods Design a DNA sequence with a small hairpin structure,PCR amplification a siRNA expression cassettes,cloning vector to construct recombinants in psi Lent Gene TM into E. coli DH5α strain,extract plasmid 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 successfully constructed. Conclusion The successful construction of STAT3 targeted RNA interference recombinant,provides some new methods for gene therapy of cancer research at the molecular level,cellular level and the overall level of.