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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。
OBJECTIVE: To construct a multi-epitope eukaryotic expression vector of human survivin and express it in human dendritic cells (DCs). METHODS: Four HLA-A2 restricted CD8 + CTL epitopes and one CD4 + Th epitope recombinant cDNA fragment containing four survivin genes were synthesized and cloned into pBluescript II SK (+) vector. After sequencing, the target gene was ligated To eukaryotic expression vector pIRESneo3.0, the recombinant eukaryotic expression plasmid pPIRESneo3.0-survivin (4) / Th. They were transfected into human DC and screened for stable expression. Results: A multi-epitope eukaryotic expression vector containing survivin was constructed and successfully transfected into human DCs. CONCLUSION: The multi-epitope eukaryotic expression vector containing survivin was successfully constructed and stably expressed in human DCs, which laid the foundation for the further study of multi-epitope tumor vaccine.