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Engineered GFP as a vital reporter in plants
1Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
Current Biology : CB
|March 1, 1996
Summary
Modified green-fluorescent protein (GFP) enhances expression and brightness in plants. This versatile reporter aids in visualizing gene expression, protein localization, and signal transduction in living plant cells and transgenic plants.
Area of Science:
- Molecular Biology
- Plant Biotechnology
Background:
- The green-fluorescent protein (GFP) from jellyfish is a reporter in various organisms.
- Enhanced expression and fluorescence intensity are needed for GFP in intact plants.
Purpose of the Study:
- To develop a modified GFP with improved expression and sensitivity for plant applications.
- To demonstrate its utility in living plant cells and transgenic plants.
Main Methods:
- Re-engineered GFP gene sequence using favored human protein codons.
- Introduced a chromophore mutation (Ser65Thr).
- Tested in maize leaf cells and various transient expression systems.
Main Results:
- Modified GFP showed 20-fold higher expression in maize leaf cells.
- Achieved over 100-fold brighter fluorescence signals and faster chromophore formation.
- Enabled detection of weak promoters, protein targeting, and signal transduction pathways.
Conclusions:
- Modified GFP is a sensitive, economical tool for visualizing promoter activity and protein localization in plants.
- Suitable for measuring dynamic responses in signal transduction and transfection efficiency.
- Codon optimization may enable high expression in evolutionarily distant species.