Non-destructive monitoring of rpoS promoter activity as stress marker for evaluating cellular physiological status

被引:14
作者
Funabashi, H
Haruyama, T
Mie, M
Yanagida, Y
Kobatake, E
Aizawa, M
机构
[1] Tokyo Inst Technol, Dept Biol Informat, Grad Sch Biosci & Biotechnol, Midori Ku, Yokohama, Kanagawa 2268501, Japan
[2] Kyushu Inst Technol, Grad Sch Life Sci & Syst Engn, Dept Biol Funct & Engn, Wakamatsu Ku, Fukuoka 8080196, Japan
关键词
bioprocess monitoring; cellular physiological status; (p)ppGpp; rpoS promoter; stress response;
D O I
10.1016/S0168-1656(01)00446-1
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
To monitor the extent of cellular physiological stress, the activity of the rpoS promoter was evaluated as a marker of the stress pathway. A reporter plasmid was constructed by inserting the GFPuv gene under the rpoS promoter and used to transform Escherichia coli cells. The fluorescence of the GFPuv protein was measured in intact cells in a non-destructive manner. The physiological status of the cells could be conveniently monitored using the rpoS-GFPuv reporter gene with respect to the cellular growth phase and to elevated ethanol and NaCl concentrations as two examples of environmental stress factors. Comparison of the response of different E. coli strains demonstrated an essential role of the rclA gene in the induction of the rpoS-GFPuv reporter gene. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:85 / 93
页数:9
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