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VIDEO CAPTURE OF GREEN FLUORESCENT PROTEIN REPORTING IN VIVO, REAL TIME GENE EXPRESSION.   M. S. Manak, A-L. Paul, P. C. Sehnke, and R. J. Ferl. Department of Horticultural Sciences and the Biotechnology Program, University of Florida, Gainesville FL 32611-0690.

Green fluorescent protein (GFP) is a reporter gene that provides the unique ability to collect in vivo data on gene expression patterns in a non-destructive manner. When coupled to a stress inducible promoter, such as that of Arabidopsis Alcohol dehydrogenase (Adh) gene, GFP can be used to monitor a plants response to stress in real time.  The in vivo stress response can be captured with digital video imaging equipment to detect the wavelength of light being admitted by GFP.  Current data captured with this method include detailed digital footage of whole Arabidopsis plants showing stress induced gene expression patterns over a 40day period.  The positive control was transgenic Arabidopsis with a CaMV 35S promoter/ GFP fusion which causes expression through out Arabidopsis from five day old seedlings to mature seven week old plants. The negative control was transgenic Arabidopsis with a promoterless GFP construct, which exhibits no GFP expression. Arabidopsis transformed with an Adh promoter/ GFP fusion construct show specific expression patterns unique to each environmental stress applied, hypoxia, cold, drought, ABA. This stress response gene/ GFP reporter gene expression system coupled with a digital camera data collection system provides the opportunity to observe Arabidopsis gene responses, in real time and via telemetry, to stresses encountered as a result of space flight missions.  In a larger sense, this technology would allow ground-based scientists to observe gene expression patterns in orbital, lunar, Martian, or other extraterrestrial environments, without the need for crew time or sample return.

(Supported by NASA: NAG10-0145.)