Related Experiment Videos
Optimized codon usage and chromophore mutations provide enhanced sensitivity with the green fluorescent protein
1Cell Biology Group, CLONTECH Laboratories Inc., Palo Alto, CA 94303-4230, USA.
Nucleic Acids Research
|November 15, 1996
Summary
Researchers developed an enhanced green fluorescent protein (GFP) with improved brightness and expression. This advanced reporter protein offers greater sensitivity for monitoring gene expression and protein localization in various biological systems.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Wild-type green fluorescent protein (GFP) from Aequorea victoria is a widely used reporter but has limitations.
- Suboptimal characteristics include low fluorescence intensity, slow fluorescence development, complex chromophore photoisomerization, and poor expression in higher eukaryotes.
Purpose of the Study:
- To engineer an enhanced GFP (EGFP) with improved fluorescence intensity and expression levels.
- To overcome the limitations of wild-type GFP for broader applications in biological research.
Main Methods:
- Combined a GFP mutant with a higher extinction coefficient for 488 nm excitation.
- Re-engineered the GFP gene sequence using codons preferred in highly expressed human proteins.
Main Results:
- The resulting enhanced GFP exhibits significantly improved fluorescence intensity.
- Demonstrated higher expression levels in various biological systems.
- The enhanced GFP provides greater sensitivity for detecting biological processes.
Conclusions:
- The engineered enhanced GFP offers superior performance compared to wild-type GFP.
- This improved reporter protein enhances the sensitivity and reliability of gene expression and protein localization studies.
- The enhanced GFP is a valuable tool for diverse applications in cell and molecular biology.