摘要
目的:将β1整合素-绿色荧光蛋白(GFP)融合基因转染兔角膜上皮细胞并检测其对各细胞外基质蛋白的黏附能力。方法:构建β1整合素-GFP融合基因真核细胞重组表达质粒,并将其转染至兔角膜上皮细胞。利用RT-PCR、Western印迹和荧光显微镜观察融合基因表达,同时检测转染细胞的增殖情况及对各细胞外基质蛋白的黏附能力。结果:成功将β1整合素-GFP融合基因转染兔角膜上皮细胞并使其过表达;转染β1整合素对角膜上皮细胞增殖无明显影响,但使细胞对各细胞外基质蛋白的黏附能力明显增高。结论:建立了功能性β1整合素-GFP过表达的角膜上皮细胞模型,为研究β1整合素基因在角膜上皮细胞的功能及角膜细胞移植奠定基础。
Objective ：To transfect rabbit corneal epithelial （RCE） cells with β1-integrin-green fluorescent protein （GFP） fusion gene, and to detect the adhesive ability of transfected cells to extracellular matrix （ECM） protein, Methods：Eukaryotic expression vector encoding β1- integrin-GFP fusion DNA was constructed and transfected into RCE cells. The expressions of β1-integrin-GFP fusion gene was detected by reverse transcription polymerase chain reaction and Western blotting and observed under fluorescence microscope, The proliferation and the adhesive ability to extracellular matrix of RCE cells were determined. Results：The β1-integrin-GFP fusion gene was transfected into RCE cells successfully and overexpressed. The transfection had no effect on the proliferation of RCE cells, but it significantly promoted the adhesive ability to ECM protein. Concluslon：The functional RCE cells overexpressing β1-integrin-GFP fusion protein are established,and they can be used for the further biological study of the β1-integrin gene in RCE cells.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2007年第5期523-525,528,共4页
Journal of China Medical University
基金
教育部留学回国人员科研启动基金资助项目(教外司留[2005]383号)
关键词
角膜上皮细胞
Β1整合素
绿色荧光蛋白
基因转染
corneal epithelial cells
β1-integrin
green fluorescent protein
gene transfection
