Green Fluorescent Protein (GFP)

Leila D Hebshi, CLONTECH Laboratories Inc., Palo Alto, California, USA
Brigitte M Angres, CLONTECH Laboratories Inc., Palo Alto, California, USA
Xianqiang L Li, Genemed Biotechnologies Inc., San Francisco, California, USA
Steven R Kain, Agilent Technologies, Santa Clara, California, USA

Published online: July 2007

DOI: 10.1002/9780470015902.a0002663.pub2

Green fluorescent protein (GFP), a fluorescent molecule, found in the jellyfish Aequorea victoria, is a useful reporter molecule for in vitro and whole organism gene expression studies. Several modifications have produced an important class of reporter molecules with superior monitoring capabilities.

Keywords: luminescence; fluorescence; fluorescent proteins; gene therapy; reporter gene; reef coral

Figure 1. Purified green fluorescent protein (GFP).
Figure 2. Structure of green fluorescent protein.
Figure 3. (a) Excitation and (b) emission spectra of EGFP and its colour variants. Data provided by Dr David Piston.
Figure 4. HeLa cells expressing ECFP localized to the endoplasmic reticulum (ER) and EYFP localized to the Golgi apparatus. Images were taken with CCD imaging, then pseudocoloured and superimposed.
Figure 5. Transgenic mice expressing GFP. Photograph provided by Dr Masaru Okabe.
close
 References
    Andersen JB, Sternberg C, Poulsen LK et al. (1998) New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria. Applied and Environmental Microbiology 64: 2240–2246.
    Bokman SH and Ward WW (1981) Renaturation of Aequorea green- fluorescent protein. Biochemical and Biophysical Research Communications 101: 1372–1380.
    book Conn PM (ed.) (1999) Green Fluorescent Protein Methods in Enzymology, 302 San Diego, CA: Academic Press.
    Ellenberg J, Lippincott-Schwartz J and Presley JF (1998) Two-color green fluorescent protein time-lapse imaging. BioTechniques 25: 838–842, 844–846.
    book Fang Y, Dery O, Haugwitz M, Turpin P and Kain SR (2006) "Practical considerations for use of reef coral fluorescent proteins in mammalian cells: applications in fluorescence microscopy and flow cytometry". In: Kain S (eds) Green Fluorescent Protein: Properties, Applications, and Protocols, 2nd edn. New York: Wiley-Liss.
    Ghosh I, Hamilton AD and Regan L (2000) Antiparallel leucine zipper-directed protein reassembly: application to green fluorescent protein. Journal of the American Chemical Society 122: 5658–5659.
    Heim R, Prasher DC and Tsien RY (1994) Wavelength mutations and posttranslational autoxidation of green fluorescent protein. Proceedings of the National Academy of Sciences of the USA 91: 12501–12504.
    Heim R and Tsien RY (1996) Engineering green fluorescent protein for improved brightness, longer wavelengths and fluorescence resonance energy transfer. Current Biology 6: 178–182.
    Kain SR (1999) Green fluorescent protein (GFP): applications in cell based assays for drug discovery. Drug Discovery Today 4: 304–312.
    Levy J, Muldoon RR, Mazo I, Kain SR and Link CJ (1999) In vivo retroviral transduction and expression of green fluorescent protein. Methods in Enzymology 302: 358–369.
    Li X, Zhao X, Fang Y et al. (1998) Generation of destabilized enhanced green fluorescent protein as a transcription reporter. Journal of Biological Chemistry 273: 34970–34975.
    Ludin B and Matus A (1998) GIP illuminates the cytoskeleton. Trends in Cell Biology 8: 72–77.
    book Ludin B and Matus A (1999) GIP in Motion (CD-ROM). Oxford: Elsevier, in association with Trends in Cell Biology.
    Mahajan NP, Linder K, Berry G et al. (1998) Bcl-2 and Bax interactions in mitochondria probed with green fluorescent protein and fluorescence resonance energy transfer. Nature Biotechnology 16: 547.
    book Magliery TJ and Regan L (2006) "Reassembled GFP: detecting protein–protein interactions and protein expression patterns". In: Chalfie M, and Kain S (eds) Green Fluorescent Protein: Properties, Applications, and Protocols, 2nd edn. New York: Wiley-Liss.
    Matz MV, Fradkov AF, Labas YA et al. (1999) Fluorescent proteins from nonbioluminescent Anthozoa species. Nature Biotechnology 17: 969–973.
    Miyawaki A, Llopis J, Heim R et al. (1997) Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin [Letter]. Nature 388: 94.
    Ormö M, Cubitt AB, Kallio K et al. (1996) Crystal structure of the Aequorea victoria green fluorescent protein. Science 273: 1392–1395.
    Patterson GH, Knobel SM, Sharif WD, Kain SR and Piston DW (1997) Use of the green fluorescent protein and its mutants in quantitative fluorescence microscopy. Biophysical Journal 73: 2782–2790.
    Pawliuk R, Bachelot T, Raftopoulos H et al. (1998) Retroviral vectors aimed at the gene therapy of human beta-globin gene disorders. Annals of the New York Academy of Sciences 850: 151–162.
    Prasher DC, Eckenrode VK, Ward WW, Prendergast FG and Cormier MJ (1992) Primary structure of the Aequorea victoria green fluorescent protein. Gene 111: 229–233.
    Shimomura O, Johnson FH and Saiga Y (1962) Extraction, purification and properties of aequorin, a bioluminescent protein from the luminous hydromedusan, Aequorea. Journal of Cellular and Comparative Physiology 59: 223–239.
    Yang F, Moss LG and Phillips GN (1996) The molecular structure of green fluorescent protein. Nature Biotechnology 14: 1246–1251.
 Further Reading
    book Chalfie M, and Kain S (eds) (2006) Green Fluorescent Protein: Properties, Applications, and Protocols, 2nd edn. New York: Wiley-Liss.
    Gerdes H-H and Kaether C (1996) Green fluorescent protein: applications in cell biology. FEBS Letters 389: 44–47.
    Misteli T and Spector DL (1997) Applications of the green fluorescent protein in cell biology and biotechnology. Nature Biotechnology 15: 961–964.
    book Sullivan K and Kay SA (eds) (1999) "GFP biofluorescence: imaging gene expression and protein dynamics in living cells". In: Methods in Cell Biology, vol 58. San Diego, CA: Academic Press.

Related Sub-Topics:

Protein Structure | Transcription | Techniques and Tools in Molecular Biology | Biochemical and Biophysical Techniques | Proteins: Structure, Function, Metabolism
Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

Article Title: Green Fluorescent Protein (GFP)
 
How to Cite close
Hebshi, Leila D, Angres, Brigitte M, Li, Xianqiang L, and Kain, Steven R(Jul 2007) Green Fluorescent Protein (GFP). In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0002663.pub2]