中国肺癌杂志2001，Vol.4Issue(1)：20-24,5.
利用GFP逆转录病毒标记肿瘤与血管内皮细胞
Using GFP retrovirus to label tumor cells and vascular endothelia cells
谢匡成 1徐萍 1顾青 1刘文文 1王丰 1田毓华 1陈霞芳 1李川源 2黄倩1
作者信息
- 1. 上海市第一人民医院中心实验室
- 2. Duke University Medical Center
- 折叠
摘要
Abstract
Objective To prepare retrovirus which carry GFP gene and are able to label living cells simply and rapidly. Methods The recombinant retroviral vector pLNCX-GFP was constructed by inserting 780?bp GFP cDNA fragment into the MCS site of retroviral plasmid pLNCX. Both ecotropic packaging cell line ΦX-Eco and amphotropic packaging cell line ΦX-Ampho and PA317 were transfected by pLNCX-GFP with liposome. The supernate collected from transfected packaging cells was used to infect a variety of tumor cell lines and vascular endothelia cell lines. Results When packaging cells were transfected by retroviral vector pLNCX-GFP, the GFP expression could be observed in 25％～40％ of cells and GFP retrovirus then could be detected, however G418 resistant clones showed more stable GFP expression and higher retrovirus titer. The GFP retrovirus from different packaging cell line showed variant ability to infect tumor cell lines and vascular endothelia cell lines and the tumor cells infected by GFP retrovirus showed stable GFP expression in vitro. GFP transduced tumor cells could grow in syngenic animal and continue expressing GFP. Conclusion Using GFP retrovirus to label target cells represent an important advantage over conventional plasmid because they can efficiently transfer GFP gene into target cells and GFP can be stably expressed in target cells no matter in vitro or in vivo.关键词
绿色荧光蛋白（GFP） 逆转录病毒 基因转移分类
医药卫生引用本文复制引用
谢匡成,徐萍,顾青,刘文文,王丰,田毓华,陈霞芳,李川源,黄倩..利用GFP逆转录病毒标记肿瘤与血管内皮细胞[J].中国肺癌杂志,2001,4(1):20-24,5.
