Arabidopsis 2010: The Arabidopsis Localizome

为了最终破译拟南芥基因组中每个基因的功能，生成高分辨率的基因表达模式和相应基因产物的亚细胞定位图是至关重要的。报告基因之间的翻译融合，如GFP，与其染色体背景中的感兴趣基因之间的融合，提供了关于该基因表达模式的最准确信息。在拟南芥中缺乏有效的同源重组系统的情况下，大的拟南芥人工染色体可以在大肠杆菌中通过同源重组精确地标记荧光蛋白，然后转移到拟南芥基因组中。这个项目的主要目的是检验这种细菌重组系统在大规模基因功能分析中的效率和有效性。将评估获得的信息的准确性以及该系统的实用性，为具有良好特征的表达模式以及明确定义的突变表型的基因产生GFP标记的拟南芥品系。我们将考察这一过程中每一步的效率以及对全基因组水平的“可伸缩性”。最后，将研究使用额外的报告基因标签的效用，例如基于GAL4的那些标签，探索使用产生的标签克隆来赋予任何感兴趣的基因特定的明确定义的表达模式的潜力，并测试特定表达模式的功能相关性。最后，将调查这项技术对其他植物物种的可转移性。广泛的影响：在该项目中产生的所有生物材料将在测试后提供给拟南芥生物资源中心的研究社区。这些资源的可用性将通过拟南芥新闻集团(http://www.bio.net/biomail/listinfo/arab-gen).宣布基因清单、标签的位置和项目进展将发布在项目专用网页上。重要的是，该项目的长期潜在影响是生产必要的工具来生成拟南芥“定位体”，即所有拟南芥基因的高分辨率表达图谱。该项目还将提供一个极好的机会，吸引和向高中和本科生介绍功能基因组学领域，特别是那些来自代表性不足群体的学生，特别是拉美裔学生。

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.