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目的:构建FHL2基因的siRNA真核表达载体,观察其对FHL2蛋白表达的影响。方法:利用RNAi干扰技术,设计并合成了两条针对FHL2基因的siRNA,将其克隆到siR-NA表达载体pSliencer 2.1-U6 neo上。将重组质粒和带FLAG标签的FHL2共转染293T人胚肾细胞,通过Western blot实验检验RNAi干扰效应。结果:经过酶切和测序证明,成功构建了FHL2 siRNA真核表达载体。通过Western blot证明,构建的siRNA能有效抑制FHL2基因的表达。结论:成功地构建了FHL2基因的siRNA真核表达载体,转染细胞后能有效地抑制FHL2基因的表达。
Objective: To construct siRNA eukaryotic expression vector of FHL2 gene and observe its effect on FHL2 protein expression. Methods: Two siRNAs targeting FHL2 gene were designed and synthesized by RNA interference technique. The siRNAs were cloned into the siR-NA expression vector pSliencer 2.1-U6 neo. 293T human embryonic kidney cells were co-transfected with the recombinant plasmid and FLAG-tagged FHL2, and the RNAi interference effect was examined by Western blot. Results: After digestion and sequencing, we successfully constructed the eukaryotic expression vector of FHL2 siRNA. Western blot confirmed that the constructed siRNA can effectively inhibit the expression of FHL2 gene. Conclusion: The eukaryotic expression vector of siRNA targeting FHL2 gene was successfully constructed and the expression of FHL2 gene was inhibited after transfection.