Saito M, Chikazawa T, Matsuoka H, Nishizawa Y, Shibuya N
Department of Biotechnology, Faculty of Technology, Tokyo University of Agriculture and Technology, Koganei, Japan.
J Biotechnol. 2000 Jan 21;76(2-3):227-32. doi: 10.1016/s0168-1656(99)00198-4.
In order to analyze intracellular signal transduction, we investigated the mechanism of chemical elicitor action by single-cell transient assay using green fluorescent protein (GFP) as a reporter gene. When the elicitor was applied from outside the cell into which the chitinase promoter and GFP reporter were introduced beforehand, fluorescence emission of GFP was observed. In contrast, when the elicitor was introduced in the cell to let the elicitor act from inside, no emission was observed. Addition of further elicitor from outside, however, did cause GFP emission. Therefore, it is clear that the elicitor does not act after entering the cell but that its signal is transduced into the cell via the cell membrane.
为了分析细胞内信号转导,我们使用绿色荧光蛋白(GFP)作为报告基因,通过单细胞瞬时分析来研究化学诱导子的作用机制。当将诱导子从预先导入了几丁质酶启动子和GFP报告基因的细胞外部施加时,观察到了GFP的荧光发射。相反,当将诱导子导入细胞内使其从内部起作用时,则未观察到发射。然而,从外部添加更多的诱导子确实会导致GFP发射。因此,很明显诱导子进入细胞后并不起作用,而是其信号通过细胞膜转导进入细胞。