Arabidopsis 2010: The Arabidopsis Localizome

拟南芥 2010：拟南芥定位组

• 批准号: 0820755
• 负责人: Jose Alonso
• 金额: $ 25万
• 依托单位: North Carolina State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-12-01 至 2011-11-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0820755&HistoricalAwards=false
• 关键词: Arabidopsis; 2010; Localizome

Towards the ultimate goal of deciphering the function of every gene in the Arabidopsis genome, it is paramount to generate high-resolution spatial and temporal maps of gene expression patterns and sub-cellular localization of the corresponding gene products. Translational fusions between a reporter gene, such as GFP, and a gene of interest in its chromosomal context provide the most accurate information on that gene's expression pattern. In the absence of efficient homologous recombination system in Arabidopsis, large Arabidopsis artificial chromosomes can be precisely tagged with fluorescence proteins by homologous recombination in E. coli and then transferred to the Arabidopsis genome. The main objective of this project is to examine the efficiency and effectiveness of this bacterial recombination system for large-scale gene function analysis in Arabidopsis. The accuracy of the obtained information and, therefore, the utility of the system will be evaluated generating GFP-tagged Arabidopsis lines for genes with well-characterized expression patterns, as well as with well-defined mutant phenotypes. The efficiency of each step in the process and the "scalability" to a whole-genome level will be examined. Finally, the utility of using additional reporter gene tags such as those based on GAL4 will be investigated, exploring the potential of employing the generated tagged clones to confer specific well-defined expression patterns to any gene of interest and to test the functional relevance of a particular expression pattern. Finally, the transferability of this technology to other plant species will be investigated.Broader Impacts: All biological materials generated in this project will be made available, upon testing, to the research community from the Arabidopsis Biological Resource Center. The availability of these resources will be announced through the Arabidopsis News Group (http://www.bio.net/biomail/listinfo/arab-gen). List of genes, position of the tags, and project progress will be posted in a project-dedicated webpage. Importantly, the long-term potential impact of this project is the production of essential tools for generating the Arabidopsis "localizome", high-resolution expression maps of all Arabidopsis genes. This project will also provide an excellent opportunity for attracting and introducing the field of functional genomics to high school and undergraduate students, particularly those from underrepresented groups and especially Hispanics.

为了实现破译拟南芥基因组中每个基因的功能的最终目标，生成基因表达模式的高分辨率时空图和相应基因产物的亚细胞定位是至关重要的。报告基因（如GFP）与目的基因在其染色体背景下的翻译融合提供了关于该基因表达模式的最准确信息。在拟南芥缺乏有效的同源重组系统的情况下，通过在E. coli中，然后转移到拟南芥基因组中。本项目的主要目的是检验这种细菌重组系统在拟南芥大规模基因功能分析中的效率和有效性。所获得的信息的准确性，因此，该系统的效用将进行评估，产生GFP标记的拟南芥品系的基因具有良好的特征表达模式，以及明确定义的突变体表型。将检查该过程中每个步骤的效率和全基因组水平的“可扩展性”。最后，将研究使用额外报告基因标签（例如基于GAL 4的标签）的实用性，探索使用生成的标记克隆来为任何感兴趣的基因赋予特定的明确表达模式并测试特定表达模式的功能相关性的潜力。模式。最后，还将研究该技术在其他植物物种上的可移植性。更广泛的影响：在该项目中产生的所有生物材料将在测试后从拟南芥生物资源中心提供给研究界。这些资源的可用性将通过拟南芥新闻组（http：//www.bio.net/biomail/listinfo/arab-gen）公布。基因列表、标签位置和项目进度将在项目专用网页上发布。重要的是，该项目的长期潜在影响是产生拟南芥“localizome”，所有拟南芥基因的高分辨率表达图谱的基本工具。该项目还将提供一个极好的机会，吸引和介绍功能基因组学领域的高中和本科生，特别是那些来自代表性不足的群体，特别是西班牙裔。