人Survivin基因真核表达载体的构建及在肝癌细胞中的表达

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

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。

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. Coneluslons： The recombinant plasmid pEGFP-Survivin was successfully constructed.