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Properties and Applications of EGFP, Enhanced Color Variants of GFP, and Unstable Derivatives of GFP

Published online by Cambridge University Press:  02 July 2020

P.A. Kitts ,
X. Li ,
D.W. Piston ,
R. Chervenak  and
S.R. Kain
P.A. Kitts
Affiliation:
CLONTECH Laboratories, Inc., Palo Alto, CA94303
X. Li
Affiliation:
CLONTECH Laboratories, Inc., Palo Alto, CA94303
D.W. Piston
Affiliation:
Department of Molecular Physiology and Biophysics, Vanderbilt University Medical School, Nashville, TN37232
R. Chervenak
Affiliation:
Department of Microbiology and Immunology, Louisiana State University Medical Center, Shreveport, LA, 71130
S.R. Kain
Affiliation:
CLONTECH Laboratories, Inc., Palo Alto, CA94303
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Extract

The green fluorescent protein (GFP) has great potential as a tool for biologists because it can be used as an in vivo real time reporter of protein localization and gene expression in a variety of experimental systems. Wild type GFP, however, has several undesirable properties including low brightness, a significant lag in the development of fluorescence, complex photoisomerization, inefficient protein folding at 37°C, and poor expression in several species. To improve upon these qualities, we have combined an ultra-bright variant of GFP, GFPmutl, with a synthetic gene sequence containing codons preferentially found in highly expressed human proteins. The combination of improved fluorescence intensity and higher expression levels yields an enhanced variant (EGFP) (Table 1) that greatly increases the sensitivity of this reporter.

The natural green emission of GFP can conveniently be monitored by optics designed to detect fluorescein. There are, however, many potential applications for GFP that require additional emission colors.

Type
Detection and Application of Green (and other Colored) Fluorescent Proteins
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 1000 - 1001
Copyright
Copyright © Microscopy Society of America

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References

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