Engineered GFP as a vital reporter in plants
Key findings
sGFP with optimized human codons achieved 20-fold higher expression than native jellyfish GFP in maize cells. Combined with S65T mutation, sGFP(S65T) produced >100-fold brighter fluorescence under 490nm excitation and enabled detection within 3-4 hours versus 15-16 hours for native GFP.
sGFP(S65T) detected weak Arabidopsis CAB2 promoter activity in maize cells (previously undetectable with native GFP) and enabled light-responsive expression. The construct worked across multiple plant systems including tobacco protoplasts (>80% transfection efficiency), Arabidopsis tissues, and onion epidermal cells.
Transgenic tobacco plants expressing 35SC4PPDK-sGFP(S65T) showed clear yellow fluorescence in all cell types, enabling rapid visual identification without drug selection or PCR. Nuclear and plastid targeting via NLS and transit peptide fusions demonstrated precise subcellular localization in living cells.