人annexin A2基因真核表达的构建及活性

为了构建人Annexin A2基因真核表达载体。通过TRIzol法提取Hela宫颈癌细胞中总RNA,采用RT-PCR技术(逆转录-聚合酶链反应)扩增Annexin A2的基因片段,并成功构建成VR1012-Annexin A2-HA重组质粒。经酶切、测序检测其构建的正确性,将质粒瞬时转染到293T人胚肾细胞,使用免疫荧光和蛋白印迹(Western blot)法检测基因表达,使用流式细胞术检测转染后细胞凋亡率。结果表明实验成功构建了具有表达活性的Annexin A2真核表达质粒,为进一步研究Annexin A2的抗凋亡作用奠定了基础。

Construction and activity of human annexin A2 gene eukaryotic expression

To construct human annexin A2 gene eukaryotic expression vector. Using TRIzol method of extracting total RNA Hela cervical cancer cells, using rt-pcr amplification annexin A2 gene fragments, and successfully build a VR1012 - annexin A2 - HA recombinant plasmid, by enzyme digestion and sequencing to detect the correctness of its building, the transient transfection plasmid to 293T embryonic kidney cells, using immunofluorescence and protein imprinting (Western blot) method to detect gene expression, using flow cytometry to detect cell apoptosis rate after transfection. In this study, we successfully constructed annexin A2 eukaryotic expression plasmid with expression activity, which laid a foundation for further study on the anti-apoptotic effect of annexin A2.