稳定表达叶酸受体α的C26细胞株的建立

发布时间：2018-12-11 22:06

【摘要】：目的建立稳定表达叶酸受体α(FRα)的C26细胞,用于FRα基因疫苗的后续研究。方法采用脂质体转染法,将前期构建的重组真核表达质粒pc DNA3.1-FRα基因导入C26细胞,用G418加压进行筛选,并通过单克隆操作,建立稳定转染叶酸受体α基因的单克隆细胞株。利用反转录PCR及免疫荧光细胞化学染色检测稳定转染细胞中叶酸受体α基因的表达情况,并利用反转录PCR检测传代细胞中FRα基因表达的稳定性。结果经转染、G418筛选以及单克隆化操作,得到单克隆FRα基因稳定转染细胞株,所得单克隆稳定转染细胞株可表达FRαmRNA及蛋白,并能够在多次传代后保持表达的稳定性。结论成功构建了FRα稳定转染细胞株。
[Abstract]:Objective to establish C26 cells stably expressing folate receptor 伪 (FR 伪) for further study of FR 伪 gene vaccine. Methods Recombinant eukaryotic expression plasmid pc DNA3.1-FR 伪 was transfected into C26 cells by liposome transfection. A monoclonal cell line stably transfected with folate receptor 伪 gene was established. Reverse transcription PCR and immunofluorescence cytochemical staining were used to detect the expression of folate receptor 伪 gene in stable transfected cells, and reverse transcription PCR was used to detect the stability of FR 伪 gene expression. Results after transfection, G418 screening and monoclonal manipulation, the stable transfection cell line of FR 伪 gene was obtained, and the stable expression of FR 伪 mRNA and protein could be maintained after multiple passages. Conclusion the stable transfection cell line of FR 伪 was successfully constructed.
【作者单位】：中国药科大学生命科学与技术学院分子生物学教研室;南京中医药大学中药学一级学科;
【基金】：国家自然科学基金(81301902,81102899)
【分类号】：R392

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