The Expression of GFP Gene in Transformed and Tumor Cells
- VernacularTitle:绿色荧光蛋白GFP在转化细胞和肿瘤细胞中的表达
- Author:
Lili, YU
;
Shishu CHEN
- Publication Type:Journal Article
- Keywords:
GFP expression reporter gene tumor gene therapy
- From:Journal of Shanghai Jiaotong University(Medical Science)
2000;20(5):408-411
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo search the appropriate experimental conditions for using green fluores-cent protein (GFP) as a reporter gene in tumor gene therapy. Methods The plasmids carrying mu-tated GFP gene were transfected into the eukaryotic cells to observe transient gene expression. Thesestudies were conducted to 1. directly determine GFP expression and express stability in the COS- 7cells, 2. compare the transfection efficiency of two plasmids pcDNA3 - EGFP, pSVKa - S65T with dif-ferent promoters in different tumor cell lines, 3. determine two genes expression in a single cell usingLacZ cotransfected with GFP by FACS. ResultsFluorescence could be detected in intact viable cellsunder different sets of conditions. The expression of GFP might last two weeks or more and the ex-pressed fluorescence was stable. The transfection rate of pcDNA3 - EGFP expressed was different inthree tumor cell lines examined. But pSVK3 - GFP expressed similarly in four tumor cell lines exam-ined. FACS showed the probability of two genes entering a single cell is above >85% at the ratio 1:4.ConclusionThe above data indicate that the GFP can be visualized continuously and directly for geneexpression in living cells. GFP may also be used for quicklly selecting cells carrying a target gene.
