人Survivin基因真核表达载体的构建及在肝癌细胞中的表达

目的:研究Survivin在肝癌细胞中的表达定位。方法:用RT-PCR的方法从肝癌组织中获取Survivin基因,并克隆到真核表达载体pEGFP-N2中,获得重组质粒pEGFP-Survivin,再转染到肝癌细胞SMMC-7721中,利用载体上的GFP跟踪其细胞内定位。结果:成功克隆了Survivin基因,经DNA测序与人Survivin基因完全一致。重组质粒pEGFP-Survivin成功转染到人肝癌细胞SMMC-7721中,并观察到GFP表达于肝癌细胞的胞浆中。结论:重组质粒pEGFP-Survivin能成功构建。

Cloning and expression of human anti-apoptotic gene Survivin in hepatoma carcinoma cells

Objective:To clone and identify the expressed location of human antiapoptotic gene-Survivin in hepatic carcinoma.Methods:Survivin cDNA was obtained by using RT-PCR method with total RNA extracted from the human hepatic carcinoma,and it was subcloned into eukaryotic expression vector pEGFP-N2,and then pEGFP-Survivin was obtained.After transiently transfecting the human liver cancer cell SMMC-7721 with the recon contained Survivin fragment,confocal microscopy was applied to observe the location of transfected Survivin in liver cancer cells.Results:The Survivin gene was cloned successfully and the homology of its sequence was 100% with human Survivin gene.The recombinant plasmid pEGFP-Survivin was transfected into SMMC-7721 cells successfully and it was expressed in cytoplasm of human liver cancer cells.Conclusions:The recombinant plasmid pEGFP-Survivin was successfully constructed.