Using Transgenic Plants that Express Green Fluorescent Protein in Teaching Quantitative Experimental Skills to First Year Undergraduates

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This project develops and field-tests materials and technology that allow first-year undergraduates to perform open-ended quantitative experiments on an environmentally regulated gene expressed in model plant systems. This is accomplished by creating transgenic plants that express a mutant form of jellyfish Green Fluorescent Protein (GFP). GFP gene expression is detected by illuminating whole plants with visible blue light. The value of this gene for the undergraduate laboratory is that students can now detect gene expression in real time and quantify the level of expression. Intellectual Merit: To circumvent the problems associated with older laboratory techniques such as slow histochemical assays that produce only qualitative data, or quantitative assays that require an expensive fluorometer, this project develops a model plant system that reports the regulation of the cor15a promoter via GFP. The GFP fluorescence is measured using inexpensive digital cameras fitted with appropriate filters and cost-free image analysis software. Undergraduate students using the new techniques are expected to become knowledgeable in molecular techniques, statistical analysis of data and interpretation of results.Broader Impacts: The pedagogy used advances the goals of the National Research Council's "Biology 2010" concept by teaching "the process of science" with quantitative investigation. The materials and procedures developed can be used on a national basis. One barrier for resource-challenged undergraduate institutions, such as community colleges, is the expense of research-quality image analysis equipment. This barrier is overcome through the building and testing of prototype instruments constructed from off-the-shelf digital imaging devices (e.g., SLR-style digital cameras) with color barrier filters and blue light-emitting diode (LED) arrays. The students quantify the level of GFP gene expression by analyzing the resulting imaging files using "ImageJ," a Java image analysis program provided free of charge over the Internet by the National Institutes of Health. National dissemination is accomplished by conducting two workshops at professional meetings, making instructional materials available on the Internet, and providing transgenic seeds to instructors. The project is piloted at the home institution, with a rich diversity of declared biology majors (75% female, 17% African American, 10% of Middle Eastern descent, and 4% Hispanic).