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

该项目开发和现场测试材料和技术，允许一年级本科生对模式植物系统中表达的环境调节基因进行开放式定量实验。这是通过创造表达水母绿色荧光蛋白（GFP）突变形式的转基因植物来实现的。用可见蓝光照射整株植物，检测GFP基因表达。这种基因对于本科实验室的价值在于，学生现在可以实时检测基因表达并量化表达水平。智力优势：为了避免与旧的实验室技术相关的问题，例如缓慢的组织化学分析只能产生定性数据，或者需要昂贵的荧光仪进行定量分析，该项目开发了一个模型植物系统，通过绿色荧光蛋白报告cor15a启动子的调节。GFP荧光测量使用配备适当滤光片和免费图像分析软件的廉价数码相机。使用这些新技术的本科生应具备分子技术、数据统计分析和结果解释方面的知识。更广泛的影响：使用的教学方法通过定量调查教授“科学的过程”，推进了国家研究委员会“生物学2010”概念的目标。所开发的材料和程序可在国家基础上使用。资源匮乏的本科院校，如社区学院，面临的一个障碍是研究质量的图像分析设备的费用。这一障碍是通过构建和测试由现成的数字成像设备（例如单反式数码相机）构建的原型仪器来克服的，这些设备带有彩色屏障滤光片和蓝色发光二极管（LED）阵列。学生们通过使用“ImageJ”（一个由美国国立卫生研究院在互联网上免费提供的Java图像分析程序）分析生成的图像文件来量化GFP基因表达水平。通过在专业会议上举办两次讲习班、在互联网上提供教学材料和向教员提供转基因种子，实现了全国传播。该项目在国内机构试点，拥有丰富的生物专业多样性（75%的女性，17%的非洲裔美国人，10%的中东血统，4%的西班牙裔）。

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).