Green fluorescent protein quantification in whole plants

Area of Science:

• Biotechnology
• Plant Science
• Molecular Biology

Background:

• Accurate, non-destructive protein quantification is vital for commercial transgenic plant applications.
• Green fluorescent protein (GFP) is a promising marker for monitoring transgenic plants and recombinant protein levels in field settings.

Purpose of the Study:

• To evaluate the utility of GFP fluorescence for quantifying protein concentrations in whole plant tissues.
• To compare the effectiveness of a portable GFP meter with a laboratory-based spectrofluorometer.

Main Methods:

• Quantification of GFP fluorescence in whole plant tissues using a portable General Fluorescence Plant Meter and a laboratory-based spectrofluorometer.
• Analysis of GFP fluorescence variability across different leaf positions and over time.
• Correlation analysis between GFP fluorescence intensity and soluble protein concentration or recombinant protein levels.

Main Results:

• GFP fluorescence intensity varied significantly with leaf age and over time, with younger leaves showing higher fluorescence.
• A direct correlation was observed between GFP fluorescence intensity and both total soluble protein concentration and Bacillus thuringiensis (Bt) cry1Ac endotoxin protein levels.
• The portable GFP meter demonstrated effectiveness in quantifying GFP fluorescence in whole plants.

Conclusions:

• GFP fluorescence serves as a reliable indicator of protein concentration in transgenic plants.
• Non-destructive assays using GFP fluorescence are valuable for monitoring and quantifying recombinant proteins under field conditions.
• Leaf age and time are critical factors influencing GFP fluorescence measurements.