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目的构建稳定表达人CD244分子转基因细胞株。方法通过RT-PCR克隆人CD244基因,将人CD244基因重组入逆转录病毒表达载体pEGZ-Term。将重组逆转录病毒载体pEGZ-Term/CD244和辅助病毒载体用脂质体法共转染包装细胞293T,收集含有完整重组逆转录病毒颗粒的293T细胞培养上清感染L929细胞,筛选获得Zeocin抗性的基因转染细胞。结果成功克隆人CD244基因,并稳定表达人CD244的基因转染细胞株L929/CD244。结论本研究成功构建了稳定表达人CD244的基因转染细胞株,为研究CD244生物学功能奠定了基础。
Objective To construct a stably expressing human CD244 transgenic cell line. Methods The human CD244 gene was cloned by RT-PCR and the human CD244 gene was recombined into the retroviral vector pEGZ-Term. The recombinant retroviral vector pEGZ-Term / CD244 and the helper virus vector were co-transfected into 293T packaging cells using lipofectamine 2000. The 293T cell culture supernatant containing intact recombinant retrovirus particles was infected into L929 cells and screened for Zeocin resistance The gene transfection cells. Results The human CD244 gene was successfully cloned and the human CD244 gene was stably transfected into L929 / CD244 cells. Conclusions This study successfully constructed a gene transfected cell line stably expressing human CD244, which laid the foundation for the study of the biological function of CD244.