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目的建立稳定表达增强型绿色荧光蛋白(EGFP)的H460细胞株,优化建系方法。方法质粒转染建立表达EGFP的H460细胞株,稀释法挑选表达EGFP的阳性单克隆,荧光显微镜观察绿色荧光,Western blot检测EGFP表达,流式细胞仪分选EGFP表达阳性的细胞。结果成功建立稳定表达EGFP的H460细胞株,其EGFP阳性表达率高。结论建立稳定表达EGFP的H460细胞株,为以后建细胞系提供更简便有效的方法,H460-EGFP可以作为研究工具用于后续观察抗癌新药体内的抗癌作用。
Objective To establish an H460 cell line stably expressing enhanced green fluorescent protein (EGFP) and optimize the method of establishing a line. Methods H460 cells expressing EGFP were established by plasmid transfection. Positive clones expressing EGFP were selected by dilution method. Green fluorescence was observed by fluorescence microscopy. EGFP expression was detected by Western blot. EGFP positive cells were sorted by flow cytometry. Results H460 cells stably expressing EGFP were successfully established and the positive rate of EGFP expression was high. Conclusion The establishment of H460 cell line stably expressing EGFP will provide a more convenient and effective method for the subsequent cell line construction. H460-EGFP may be used as a research tool for the subsequent observation of anticancer effects of the new anticancer drug in vivo.