Use of the A. Victoria Green Fluorescent Protein to Study Protein Dynamics In Vivo

Jason A. Kahana1, Pam A. Silver2

1 University of California, San Diego, California, 2 Dana‐Farber Cancer Institute, Boston, Massachusetts
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Unit 5.15
DOI:  10.1002/0471142301.ns0515s14
Online Posting Date:  May, 2001
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Fluorescent molecules serve as valuable tools for the detection of numerous biochemical phenomena and have been employed for protein localization, quantitation of gene expression, detection of nucleic acids, cell sorting and determination of chemical concentrations. However, the use of such techniques generally requires significant nonphysiological perturbations to the biological system being studied; therefore, they are not always appropriate for the observation of dynamic phenomena. Green fluorescent protein (GFP), cloned from jellyfish, has been used to overcome many of these problems. It is a small, extremely stable fluorescent protein that has been successfully expressed and detected in a wide variety of organisms, both in intact form and fused to other proteins. This overview unit describes the use of this proteinaceous fluorophore for in vivo observation of cellular phenomena.

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Table of Contents

  • Overview of GFP Fluorescence
  • Utilization of GFP
  • Problems with GFP
  • Mutants of GFP
  • Literature Cited
  • Figures
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Literature Cited

Literature Cited
   Chalfie, M., Tu, Y., Euskirchen, G., Ward, W.W., and Prasher, D.C. 1994. Green fluorescent protein as a marker for gene expression. Science 263:802‐805.
   Cody, C.W., Prasher, D.C., Westler, W.M., Pendergast, F.G., and Ward, W.W. 1993. Chemical structure of the hexapeptide chromophore of the Aequorea green‐fluorescent protein. Biochemistry 32:1212‐1218.
   Delagrave, S., Hawtin, R.E., Silva, C.M., Yang, M.M., and Youvan, D.C. 1995. Red‐shifted excitation mutants of the green fluorescent protein. Bio/Technology 13:151‐154.
   Flach, J., Bossie, M., Vogel, J., Corbett, A.H., Jinks, T., Willins, D.A., and Silver, P.A. 1994. A yeast RNA‐binding protein shuttles between the nucleus and the cytoplasm. Mol. Cell. Biol. 14:8399‐8407.
   Haseloff, J. and Amos, B. 1995. GFP in plants. Trends Genet. 11:328‐329.
   Hecht, E. 1987. Optics, 2nd ed. Addison‐Wesley Publishing, Reading, Mass.
   Heim, R., Cubitt, A.B., and Tsien, R.Y. 1995. Improved green fluorescence. Nature 373:663‐664.
   Heim, R., Prasher, D.C., and Tsien, R.Y. 1994. Wavelength mutations and posttranslational autoxidation of green fluorescent protein. Proc. Natl. Acad. Sci. U.S.A. 91:12501‐12504.
   Inouye, S. and Tsuji, F.I. 1994a. Aequorea green fluorescent protein: Expression of the gene and fluorescence characteristics of the recombinant protein. FEBS Lett. 341:277‐280.
   Inouye, S. and Tsuji, F.I. 1994b. Evidence for redox forms of the Aequorea green fluorescent protein. FEBS Lett.. 351:211‐214.
   Kahana, J.A., Schnapp, B.J., and Silver, P.A. 1995. Kinetics of spindle pole body separation in budding yeast. Proc. Natl. Acad. Sci. U.S.A. 92:9707‐9711.
   Morin, J.G. and Hastings, J.W. 1971. Energy transfer in a bioluminescent system. J. Cell Physiol. 77:313‐318.
   Morise, H., Shimomura, O., Johnson, F.H., and Winant, J. 1974. Intermolecular energy transfer in the bioluminescent system of Aequorea. Biochemistry 13:2656‐2663.
   Olson, K.R., McIntosh, J.R., and Olmstead, J.B. 1995. Analysis of MAP4 function in living cells using green fluorescent protein (GFP) chimeras. J. Cell Biol. 130:639‐650.
   Pines, J. 1995. GFP in mammalian cells. Trends Genet. 11:326‐327.
   Prasher, D.C., Eckenrode, V.K., Ward, W.W., Pendergast, F.G., and Cormier, M.J. 1992. Primary structure of the Aequorea victoria green‐fluorescent protein. Gene 111:229‐233.
   Reichman, J. 1994. Handbook of Optical Filters for Fluorescence Microscopy. Chroma Technology Corporation, Brattleboro, Vt.
   Wang, S. and Hazelrigg, T. 1994. Implications for bcd mRNA localization from spatial distribution of exu protein in Drosophila oogenesis. Nature 369:400‐402.
   Ward, W.W. 1981. Properties of the coelenterate green fluorescent proteins. In Bioluminescence and Chemiluminescence: Basic Chemistry and Analytical Applications (M.A. DeLuca and W.D. McElroy, eds.) pp. 225‐234. Academic Press, San Diego.
   Ward, W.W. and Bokman, S.H. 1982. Reversible denaturation of Aequorea green fluorescent protein: Physical separation and characterization of the renatured protein. Biochemistry 21:4535‐4540.
   Ward, W.W., Cody, C.W., Hart, R.C., and Cormier, M.J. 1980. Spectrophotometric identity of the energy transfer chromophores in Renilla and Aequorea green‐fluorescent proteins. Photochem. Photobiol. 31:611‐615.
   Yocum, R.R., Hanley, S., West, R., and Ptashne, M. 1984. Use of LacZ fusions to delimit regulatory elements of the inducible divergent GAL1‐GAL10 promoter in Saccharomyces cerevisiae. Mol. Cell. Biol. 4:1985‐1998.
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