Skip to main content
SpringerLink
Log in

Fluorescent monitoring of cellular physiological status depending on the accumulation of ppGpp

  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

For evaluating the physiological status of cells, astringent response network was used. Fluorescence from intact E. coli, which has a plasmid encoding the green fluorescence protein (GFP) under the regulation of rpoS promoter, was monitored. Comparison of the response of different E. coli strains demonstrated an essential role of ppGpp in the expression of GFP, as it activated the rpoS promoter. The physiological status of intact cells, that depends on ppGpp accumulation in response to the nutritional status such as amino acid starvation, could therefore be monitored by measuring fluorescent intensity using this reporter gene.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Cashel M (1969) The control of ribonucleic acid synthesis in Escherichia coli. IV. Relevance of unusual phosphorylated compounds from amino acid-starved stringent strains. J. Biol. Chem. 244: 3133-3141.

    Google Scholar 

  • Cashel M, Gentry DR, Hernandez VJ, Vinella D (1996) The stringent response. In: Neidhardt FC, Curtiss III R, Ingraham JL, Lin ECC, Low KB, Magasanik B, Reznikoff WS, Riley M, Schaechter M, Umbarger HE, eds. Escherichia coli and Salmonella. Cellular and Molecular Biology, 2nd edn. Washington D.C.: American Society for Microbiology, pp. 1458-1496.

    Google Scholar 

  • Gill RT, Valdes JJ, Bentley WE (2000) A comparative study of global stress gene regulation in response to overexpression of recombinant proteins in Escherichia coli. Metab. Eng. 2: 178-189.

    Google Scholar 

  • Hengge-Aronis R (1996) Regulation of gene expression during entry into stationary phase. In: Neidhardt FC, Curtiss III R, Ingraham JL, Lin ECC, Low KB, Magasanik B, Reznikoff WS, Riley M, Schaechter M, Umbarger HE, eds. Escherichia coli and Salmonella. Cellular and Molecular Biology, 2nd edn. Washington D.C.: American Society for Microbiology, pp. 1497-1512.

    Google Scholar 

  • Jishage M, Ishihama A (1995) Regulation of RNA polymerase sigma subunit synthesis in Escherichia coli: intracellular levels of sigma 70 and sigma 38. J. Bacteriol. 177: 6832-6835.

    Google Scholar 

  • Jishage M, Iwata A, Ueda S, Ishihama A (1996) Regulation of RNA polymerase sigma subunit synthesis in Escherichia coli: intracellular levels of four species of sigma subunit under various growth conditions. J. Bacteriol. 178: 5447-5451.

    Google Scholar 

  • Lange R, Fischer D, Hengge-Aronis R (1995) Identification of transcriptional start sites and the role of ppGpp in the expression of rpoS, the structural gene for the sigma S subunit of RNA polymerase in Escherichia coli. J. Bacteriol. 177: 4676-4680.

    Google Scholar 

  • Loewen PC, Hu B, Strutinsky J, Sparling R (1998) Regulation in the rpoS regulon of Escherichia coli. Can. J. Microbiol. 44: 707-717.

    Google Scholar 

  • McCann MP, Kidwell JP, Matin A (1991) The putative sigma factor KatF has a central role in development of starvation-mediated general resistance in Escherichia coli. J. Bacteriol. 173: 4188-4194.

    Google Scholar 

  • Neubauer P, Ahman M, Tornkvist M, Larsson G, Enfors SO (1995) Response of guanosine tetraphosphate to glucose fluctuations in fed-batch cultivations of Escherichia coli. J. Biotechnol. 43: 195-204.

    Google Scholar 

  • Nyström T (1998) To be or not to be: the ultimate decision of the growth arrested bacterial cell. FEMS Microbiol. Rev. 21: 283-290.

    Google Scholar 

  • Pizer LI, Merlie JP (1973) Effect of serine hydroxamate on phospholipid synthesis in Escherichia coli. J. Bacteriol. 114: 980-987.

    Google Scholar 

  • Sambrook J, Fritsch EF, Hecker M (1989) Molecular Cloning: A Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.

    Google Scholar 

  • Schuster KC (2000) Monitoring the physiological status in bioprocesses on the cellular level. Adv. Biochem. Eng. Biotechnol. 66: 185-208.

    Google Scholar 

  • Schweder T, Kruger E, Xu B, Jurgen B, Blomsten G, Enfors SO, Hecker M (1999) Monitoring of genes that respond to processrelated stress in large-scale bioprocesses. Biotechnol. Bioeng. 65: 151-159.

    Google Scholar 

  • Tanaka K, Handel K, Loewen PC, Takahashi H (1997) Identification and analysis of the rpoS-dependent promoter of katE, encoding catalase HPII in Escherichia coli. Biochim. Biophys. Acta 1352: 161-166.

    Google Scholar 

  • Teich A, Meyer S, Lin HY, Andersson L, Enfors S, Neubauer P (1999) Growth rate related concentration changes of the starvation response regulators sigmaS and ppGpp in glucose-limited fed-batch and continuous cultures of Escherichia coli. Biotechnol. Prog. 15: 123-129.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama, 226-8501, Japan

    Hisakage Funabashi, Masayasu Mie, Yasuko Yanagida, Eiry Kobatake & Masuo Aizawa

Authors
  1. Hisakage Funabashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Masayasu Mie
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yasuko Yanagida
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Eiry Kobatake
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Masuo Aizawa
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Funabashi, H., Mie, M., Yanagida, Y. et al. Fluorescent monitoring of cellular physiological status depending on the accumulation of ppGpp. Biotechnology Letters 24, 269–273 (2002). https://doi.org/10.1023/A:1014032711683

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1014032711683

Search

Navigation