Loimas S, Wahlfors J, Jänne J
A.I. Virtanen Institute, University of Kuopio, Finland.
Biotechniques. 1998 Apr;24(4):614-8. doi: 10.2144/98244st01.
Green fluorescent protein (GFP) and herpes simplex virus type-I thymidine kinase (TK) are commonly used markers in gene transfer studies. The latter gene has also proven to be an effective tool in cancer "suicide" gene therapy. To facilitate rapid and reliable selection of cells expressing TK, we constructed a plasmid expressing a TK-green fluorescent protein fusion gene (TK-GFP). In this fusion gene, the expression of each component is coupled to one another, permitting accurate determination of the percentage of cells expressing TK by detecting the green fluorescence produced by GFP. Transfection of the fusion plasmid to mammalian cells revealed that the construct is fully functional, making the cells both fluorescent and sensitive to ganciclovir.
绿色荧光蛋白（GFP）和单纯疱疹病毒I型胸苷激酶（TK）是基因转移研究中常用的标志物。后一种基因也已被证明是癌症“自杀”基因治疗中的一种有效工具。为便于快速、可靠地筛选表达TK的细胞，我们构建了一种表达TK-绿色荧光蛋白融合基因（TK-GFP）的质粒。在这个融合基因中，每个组分的表达相互关联，通过检测GFP产生的绿色荧光能够准确测定表达TK的细胞百分比。将融合质粒转染至哺乳动物细胞后发现该构建体功能完全正常，使得细胞既发出荧光又对更昔洛韦敏感。
