Nucleic Acids Research, Vol 24, Issue 22 4592-4593, Copyright © 1996 by Oxford University Press
TT Yang, L Cheng and SR Kain
The green fluorescent protein (GFP) from Aequorea victoria is a versatile
reporter protein for monitoring gene expression and protein localization in
a variety of cells and organisms. Despite many early successes using this
reporter, wild type GFP is suboptimal for most applications due to low
fluorescence intensity when excited by blue light (488 nm), a significant
lag in the development of fluorescence after protein synthesis, complex
photoisomerization of the GFP chromophore and poor expression in many
higher eukaryotes. To improve upon these qualities, we have combined a
mutant of GFP with a significantly larger extinction coefficient for
excitation at 488 nm with a re-engineered GFP gene sequence containing
codons preferentially found in highly expressed human proteins. The
combination of improved fluorescence intensity and higher expression levels
yield an enhanced GFP which provides greater sensitivity in most systems.
ARTICLES
Optimized codon usage and chromophore mutations provide enhanced sensitivity with the green fluorescent protein
Cell Biology Group, CLONTECH Laboratories Inc., Palo Alto, CA 94303- 4230, USA.
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