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Applied Microbiology and Biotechnology

, Volume 70, Issue 2, pp 229–236 | Cite as

A rapid reporter system using GFP as a reporter protein for identification and screening of synthetic stationary-phase promoters in Escherichia coli

  • G. MikschEmail author
  • F. Bettenworth
  • K. Friehs
  • E. Flaschel
  • A. Saalbach
  • T. W. Nattkemper
Applied Genetics and Molecular Biotechnology

Abstract

To develop a rapid reporter system for the screening of stationary-phase promoters in Escherichia coli, the expression pattern of the green fluorescent protein (GFP) during bacterial cultivation was compared with that of the commonly used β-galactosidase. Using GFP with enhanced fluorescence, the expression pattern of both reporter systems GFP and β-galactosidase were similar and showed a typical induction of gene activity of the reporter genes, i.e. increase of expression at the transition from exponential to stationary phase. The expression was affected by the culture medium, i.e. in contrast to the complex medium (LB medium), the stationary-phase specific induction was only observed in synthetic medium (M9) when amino acids were added, whereas there was generally no induction in MOPS medium. To develop a rapid screening method on agar plates for stationary-phase promoters, a photographic approach was used, continued with computational image treatment. A screening method is presented which enables an on-line monitoring of gene activity.

Keywords

Green Fluorescent Protein Reporter System Muffler Hough Transformation Promoter Probe Vector 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

Financial support of the Deutsche Forschungsgemeinschaft (FL 292/3-2) is gratefully acknowledged. F.B. is grateful to the Max-Buchner-Stiftung for receipt of a scholarship.

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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • G. Miksch
    • 1
    Email author
  • F. Bettenworth
    • 1
  • K. Friehs
    • 1
  • E. Flaschel
    • 1
  • A. Saalbach
    • 2
  • T. W. Nattkemper
    • 2
  1. 1.Lehrstuhl für Fermentationstechnik, Technische FakultätUniversität BielefeldBielefeldGermany
  2. 2.Lehrstuhl für Angewandte Neuroinformatik, Technische FakultätUniversität BielefeldBielefeldGermany

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