Taguchi Kazuyuki, Tanaka Yoshiharu, Imaeda Takao, Hirai Masana, Mohri Shino, Yamada Masato, Inoue Yuzo
Fuji Electric Advanced Technology Co., Ltd., 1 Fuji-chou, Hino City, Tokyo 191-8502, Japan.
Environ Sci. 2004;11(5):293-302.
The umu-lux test is a genotoxicity test using the two genetically modified S. typhmurium TA1535 strains (TL210 and TL210ctl) transformed with the luxCDABE (luciferase gene and fatty acid reductase genes) of Vibrio fischeri as a reporter gene. The TL210 strain detects genotoxicants and the TL210ctl strain detects cytotoxicants. In order to develop a highly sensitive, simple and rapid genotoxicity detection system, we constructed a biosensor using these immobilized strains. The biosensor consists of two immobilized microbial membranes, a sample vessel and photodetectors, and the genotoxicity detection system consists of the biosensor, an isothermal box, a photodetector and an air pump. The total measurement time for genotoxicants using this detection system is about 4 h. When 2% (v/v) DMSO was used as a control, the TL210 strain was not emitting light while the TL210ctl strain was. When 0.3 mg/l 4NQO was used as a genotoxicant, TL210 strain and TL210ctl strain were both emitting light. When HgCl2 was used as a cytotoxicant, neither the TL210 strain nor the TL210ctl strain were emitting light. Therefore, the false negative prevention function of a biosensor using the TL210ctl strain has been checked. These results show that our proposed system can correctly detect genotoxicants.
umu-lux试验是一种遗传毒性试验,使用两种经基因改造的鼠伤寒沙门氏菌TA1535菌株(TL210和TL210ctl),它们用费氏弧菌的luxCDABE(荧光素酶基因和脂肪酸还原酶基因)作为报告基因进行了转化。TL210菌株检测遗传毒性剂,TL210ctl菌株检测细胞毒性剂。为了开发一种高度灵敏、简单快速的遗传毒性检测系统,我们使用这些固定化菌株构建了一种生物传感器。该生物传感器由两个固定化微生物膜、一个样品容器和光电探测器组成,遗传毒性检测系统由生物传感器、一个恒温箱、一个光电探测器和一个气泵组成。使用该检测系统检测遗传毒性剂的总测量时间约为4小时。当使用2%(v/v)二甲基亚砜作为对照时,TL210菌株不发光,而TL210ctl菌株发光。当使用0.3mg/l 4NQO作为遗传毒性剂时,TL210菌株和TL210ctl菌株都发光。当使用HgCl2作为细胞毒性剂时,TL210菌株和TL210ctl菌株都不发光。因此,已经检验了使用TL210ctl菌株的生物传感器的假阴性预防功能。这些结果表明,我们提出的系统能够正确检测遗传毒性剂。