Non-destructive monitoring of rpoS promoter activity as stress marker for evaluating cellular physiological status.
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 relA gene in the induction of the rpoS-GFPuv reporter gene.
Funabashi
Hisakage
H
Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan.
Haruyama
Tetsuya
T
Mie
Masayasu
M
Yanagida
Yasuko
Y
Kobatake
Eiry
E
Aizawa
Masuo
M
D000438:Alcohols / Q000494:pharmacology; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics*; D015415:Biomarkers / Q000032:analysis*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000502:physiology*; D005453:Fluorescence; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D017930:Genes, Reporter / Q000235:genetics; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012808:Sigma Factor / Q000032:analysis / Q000235:genetics*; D012965:Sodium Chloride / Q000494:pharmacology; D013997:Time Factors
D000438:Alcohols; D001426:Bacterial Proteins; D015415:Biomarkers; D008164:Luminescent Proteins; D012808:Sigma Factor; C069567:sigma factor KatF protein, Bacteria; D049452:Green Fluorescent Proteins; D012965:Sodium Chloride