稳定表达egfp基因细胞的构建与克隆

Establishment of Cell Clones Expressing EGFP Stably

将增强的绿色荧光蛋白 ( enhanced green fluorescent protein,EGFP)基因插在 HCMV( hum ancytom egolovirus)启动子下游 ,构建了表达质粒 p CA13 - e G,用脂质体 L ipofectin介导分别转染 He L a细胞、Vero细胞 ,仅通过细胞传代 ,就获得了能稳定高效表达 EGFP的绿色细胞 .比较发现 ,质粒 p CA13 - e G转染后 ,产生能高效表达 EGFP的 He L a细胞其比率高于 Vero细胞 ;EGFP高效表达对 Vero细胞的毒性大于对 He L a细胞的毒性 .本研究表明 ,绿色细胞轮廓清晰 ,由于其特有的性质 ,在用于细胞的形态观察、细胞分裂等研究时会有所作为 .

It is urgent to widen and deepen the underslanding of cellular behavior. Enhanced green fluorescent protein (EGFP) gene, the most widely utilized variant of fluorescent protein (FP) gene family, was inserted into pCA13. Recombinant plasmid pCA13-eG was constructed. It was transfected into mammalian cells, HeLa and Vero, by Lipofectin. Activated by blue light, some cells emit bright, green light Green cell clones of HeLa and Vero were gotten only by passage, respectively. The transfection ratio and the aberration of the bright, green cells demonstrate that EGFP is more toxic to Vero cells than HeLa cells. In certain certain, difference of total active EGFP molecules can be used to demonstrate the different expression level of two F1-cells. The result of computer analysis shows that the molecules of EGFP are different in two F1-cells.