Survivin多表位真核表达载体的构建及其在树突状细胞中的表达

Construction of the eukaryotic expression vector encoding multi-epitope fusion protein of human survivin and its expression in dendritic cells

目的:构建人生存素(survivin)的多表位真核表达载体,并在人树突状细胞(DC)中进行表达。方法:人工合成含4个survivin的HLA-A2类限制性CD8+CTL表位和一个CD4+Th表位的重组cDNA片段,克隆到pBluescript II SK(+)载体上,测序正确后将目的基因连接到真核表达载体pIRESneo3.0中,构建的重组真核表达质粒pPIRESneo3.0-survivin(4)/Th。将其转染人DC并筛选,进行稳定表达。结果:构建了含有survivin的多表位真核表达载体,并成功地转染了人DC。结论:成功地构建了含有survivin的多表位真核表达载体,并在人DC中稳定表达,为进一步研究多表位肿瘤疫苗奠定基础。

AIM： To construct a eukaryotic expression vector encoding the multi-epitope fusion protein of human survivin,and express it in human dendritic cells.METHODS： Recombinant cDNA sequence encoding four HLA-A2-restricted CD8＋ CTL epitopes and a CD4＋ Th epitope was synthesized and cloned into pBluescript II SK（＋） vector.After confirmed by sequencing,the cDNA fragment was inserted to eukaryotic expression vector pIRESneo3.0 to generate the recombinant plasmid pPIRESneo3.0-survivin（4）/Th.The pPIRESneo3.0-survivin（4）/Th was then transfected into human dendritic cells and the transfectants were selected for stable expression.RESULTS： The eukaryotic expression vector encoding the multi-epitope fusion protein of survivin was constructed,and successfully transfected into human dendritic cells.CONCLUSION： The eukaryotic expression vector encoding the multi-epitope fusion protein of survivin has been constructed successfully,and stably expressed in human dendritic cells,which provides clues for further research on multi-epitope cancer vaccine.