Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Reporter proteins for in vivo fluorescence without oxygen

Nature Biotechnology volume 25, pages 443–445 (2007)Cite this article

Abstract

Fluorescent reporter proteins such as green fluorescent protein are valuable noninvasive molecular tools for in vivo real-time imaging of living specimens. However, their use is generally restricted to aerobic systems, as the formation of their chromophores strictly requires oxygen. Starting with blue-light photoreceptors from Bacillus subtilis and Pseudomonas putida that contain light-oxygen-voltage–sensing domains, we engineered flavin mononucleotide–based fluorescent proteins that can be used as fluorescent reporters in both aerobic and anaerobic biological systems.

Your institute does not have access to this article

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

Buy

All prices are NET prices.

Figure 1: Generation, characterization and in vivo application of anaerobically fluorescing proteins.

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  1. Matz, M.V., Lukyanov, K.A. & Lukyanov, S.A. Bioessays 24, 953–959 (2002).

    CAS  Article  Google Scholar 

  2. Chudakov, D.M., Lukyanov, S. & Lukyanov, K.A. Trends Biotechnol 23, 605–613 (2005).

    CAS  Article  Google Scholar 

  3. March, J.C., Rao, G. & Bentley, W.E. Appl. Microbiol. Biotechnol. 62, 303–315 (2003).

    CAS  Article  Google Scholar 

  4. Tsien, R.Y. Annu. Rev. Biochem. 67, 509–544 (1998).

    CAS  Article  Google Scholar 

  5. Losi, A., Polverini, E., Quest, B. & Gärtner, W. Biophys. J. 82, 2627–2634 (2002).

    CAS  Article  Google Scholar 

  6. Krauss, U., Losi, A., Gärtner, W., Jaeger, K.-E. & Eggert, T. Phys. Chem. Chem. Phys. 7, 2804–2811 (2005).

    CAS  Article  Google Scholar 

  7. Losi, A. Photochem. Photobiol. Sci. 3, 566–574 (2004).

    CAS  Article  Google Scholar 

  8. Avila-Perez, M., Hellingwerf, K.J. & Kort, R. J. Bacteriol. 188, 6411–6414 (2006).

    CAS  Article  Google Scholar 

  9. Buttani, V. et al. Photochem. Photobiol. Sci. 6, 41–49 (2007).

    CAS  Article  Google Scholar 

  10. Swartz, T.E. et al. J. Biol. Chem. 276, 36493–36500 (2001).

    CAS  Article  Google Scholar 

  11. Losi, A., Ghiraldelli, E., Jansen, S. & Gärtner, W. Photochem. Photobiol. 81, 1145–1152 (2005).

    CAS  Article  Google Scholar 

  12. Patterson, G., Day, R.N. & Piston, D. J. Cell Sci. 114, 837–838 (2001).

    CAS  PubMed  Google Scholar 

  13. Brown, J.M. Mol. Med. Today 6, 157–162 (2000).

    CAS  Article  Google Scholar 

  14. Drepper, T. et al. Biotechnol. J. 1, 777–786 (2006).

    CAS  Article  Google Scholar 

  15. Masepohl, B., Drepper, T. & Klipp, W. Nitrogen fixation in the phototrophic purple bacterium Rhodobacter capsulatus. in Genetics and Regulation of Nitrogen Fixation in Free-Living Bacteria (Klipp, W., Masepohl, B., Gallon, J.R. & Newton, W.E., eds.) 141–173 (Kluwer Academic Publisher, Dordrecht, Boston, London, 2004).

    Google Scholar 

  16. Zhang, C., Xing, X.H. & Lou, K. FEMS Microbiol. Lett. 249, 211–218 (2005).

    CAS  Article  Google Scholar 

  17. Karatani, H., Yoshizawa, S. & Hirayama, S. Photochem. Photobiol. 79, 120–125 (2004).

    CAS  Article  Google Scholar 

  18. Bettegowda, C. et al. Nat. Biotechnol. 24, 1573–1580 (2006).

    CAS  Article  Google Scholar 

Download references

Acknowledgements

The authors would like to thank Jürgen Hubbuch and Thomas Rosenbaum, Institute of Biotechnology 2, Forschungszentrum Jülich, Germany, for their assistance with confocal laser scanning microscopy. This work was partly supported by grants from the Deutsche Forschungsgemeinschaft (DFG) within the Forschergruppe FOR526 “Sensory blue light photoreceptors.”

Author information

Author notes

  1. Thomas Drepper and Thorsten Eggert: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Molecular Enzyme Technology, Heinrich-Heine-University Duesseldorf, Research Center Juelich, Stetternicher Forst, Juelich, D-52426, Germany

    Thomas Drepper, Thorsten Eggert, Franco Circolone, Achim Heck, Ulrich Krauß, Jan-Karl Guterl, Marion Wendorff & Karl-Erich Jaeger

  2. EVOcatal GmbH, Merowingerplatz 1a, Duesseldorf, D-40225, Germany

    Thorsten Eggert

  3. Department of Physics, University of Parma, viale G.P. Usberti 7/A, Parma, I-43100, Italy

    Aba Losi

  4. Max-Planck-Institute for Bioinorganic Chemistry, Stiftstraße 34-36, Mülheim an der Ruhr, D-45470, Germany

    Wolfgang Gärtner

Authors
  1. Thomas Drepper
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Thorsten Eggert
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Franco Circolone
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Achim Heck
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ulrich Krauß
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jan-Karl Guterl
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Marion Wendorff
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Aba Losi
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Wolfgang Gärtner
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Karl-Erich Jaeger
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Thomas Drepper or Karl-Erich Jaeger.

Ethics declarations

Competing interests

A patent application has been filed covering part of the results described in this manuscript (T.E., T.D., J.-K.G., A.H. & K.-E.J., patent no. DE 10 2005 048 828.5 (2005)).

Supplementary information

Supplementary Fig. 1

Structure of the Chlamydomonas reinhardtii LOV1 domain (pdb entry: 1N9L) in the dark state. (DOC 584 kb)

Supplementary Fig. 2

The complete nucleotide and amino acid sequences of BsFbFP, EcFbFP, and PpFbFP. (DOC 30 kb)

Supplementary Methods

(DOC 47 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Drepper, T., Eggert, T., Circolone, F. et al. Reporter proteins for in vivo fluorescence without oxygen. Nat Biotechnol 25, 443–445 (2007). https://doi.org/10.1038/nbt1293

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt1293

Further reading

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing