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

使用表达绿色荧光蛋白的转基因植物向一年级本科生教授定量实验技能

• 批准号: 0442771
• 负责人: Stokes Baker
• 金额: --
• 依托单位: University of Detroit Mercy
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0442771&HistoricalAwards=false
• 关键词: Using; Transgenic; Plants; Express; Green

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).

该项目开发并现场测试了材料和技术，使一年级本科生能够对模型植物系统中表达的环境调节基因进行开放式定量实验。这是通过培育表达水母绿色荧光蛋白（GFP）突变体的转基因植物来实现的。通过用可见蓝光照射整个植物来检测 GFP 基因表达。该基因对于本科生实验室的价值在于，学生现在可以实时检测基因表达并量化表达水平。智力优点：为了规避与旧实验室技术相关的问题，例如仅产生定性数据的缓慢组织化学测定，或需要昂贵的荧光计的定量测定，该项目开发了一个模型植物系统，通过 GFP 报告 cor15a 启动子的调节。 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来测量 GFP 荧光。 使用新技术的本科生预计将掌握分子技术、数据统计分析和结果解释方面的知识。 更广泛的影响：所使用的教学法通过教授定量研究的“科学过程”，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全国范围内使用。对于社区学院等资源匮乏的本科院校来说，其面临的障碍之一是研究级图像分析设备的费用。通过构建和测试由带有彩色屏障滤光片和蓝色发光二极管 (LED) 阵列的现成数字成像设备（例如单反式数码相机）构建的原型仪器，可以克服这一障碍。学生们通过使用“ImageJ”（美国国立卫生研究院通过互联网免费提供的 Java 图像分析程序）分析生成的成像文件来量化 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以及向教师提供转基因种子来完成全国传播。 该项目在母校试行，该校拥有丰富的生物学专业学生多样性（75% 为女性，17% 为非裔美国人，10% 为中东裔，4% 为西班牙裔）。