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【目的】将G250抗原cDNA片段转染入sp2/0细胞中并获得稳定表达,为G250基因疫苗研究提供实验工具。【方法】用脂质体转染法将鉴定好的重组质粒转染入sp2/0细胞中,经G418筛选得到稳定表达,采用逆转录聚合酶链式反应(RT-PCR)、间接免疫荧光、Western blot检测G250基因在sp2/0细胞中的表达。【结果】转染2周后,筛选出阳性sp2/0细胞克隆,通过RT-PCR证实有G250mRNA转录,间接免疫荧光可以见到阳性细胞,Western blot证实有G250抗原表达。【结论】建立了稳定表达G250抗原的肿瘤细胞模型,为研究基于肾癌G250基因的肿瘤基因疫苗治疗奠定了基础。
【Objective】 The G250 antigen cDNA fragment was transfected into sp2 / 0 cells and stably expressed, which provided an experimental tool for G250 gene vaccine research. 【Method】 The recombinant plasmids were transfected into sp2 / 0 cells by lipofection method. The recombinant plasmids were transfected into sp2 / 0 cells by lipofectamine. The recombinant plasmids were stably expressed by G418 screening. RT-PCR, indirect immunofluorescence, The expression of G250 gene in sp2 / 0 cells was detected by Western blot. 【Results】 Two weeks after transfection, positive sp2 / 0 cell clones were screened and G250 mRNA was confirmed by RT-PCR. Positive cells were seen by indirect immunofluorescence and G250 antigen was confirmed by Western blot. 【Conclusion】 A tumor cell model stably expressing G250 antigen was established and laid the foundation for the study of tumor gene vaccine based on G250 gene of renal cell carcinoma.