Livin和Survivin基因联合靶向siRNA重组表达载体的构建和鉴定

Construction and Identification of Multiple siRNA Recombinant Expression Vector Targeting Livin and Survivin Gene Simultaneously

目的构建Livin和Survivin联合靶向的siRNA重组表达载体,探讨用RNA干扰方法同时下调Livin和Survivin基因表达的增效作用。方法通过前期的实验,分别从靶向Livin基因和Survivin基因的siRNA序列中各挑选出1对最有效的siRNA序列,然后采用1个载体编码2个shRNA技术,将其构建到同一个真核表达载体上,转化DH5α菌株,提取质粒进行酶切鉴定和测序分析。结果经酶切鉴定和测序分析证实Livin和Survivin联合靶向的siRNA重组表达载体构建成功。结论Livin和Survivin联合靶向的siRNA重组表达载体构建成功。

Objective To construct a novel siRNA expression vector targeting Livin and Survivin simultaneously,and investigate the synergistic effect of down-regulation of Livin and Survivin.Methods In the previous experiment,one pair of the most effective siRNA sequence was chosen from Livin and Survivin siRNA sequences respectively.The two pair of different SiRNA sequences were cloned into one single vector and transformed into DH5α strains.Recombinant expression vector was identified by restriction endonuclease and sequencing analysis.Results It was confirmed by restriction endonuclease and sequencing analysis that multiple siRNA recombinant expression vector targeting Livin and Survivin genes simultaneously was constructed successfully.Conclusion The multiple siRNA recombinant expression vector targeting Livin and Survivin gene simultaneously has been constructed successfully.