Arabidopsis 2010: The Arabidopsis Localizome

为了最终实现破译拟南芥基因组中每个基因的功能，生成基因表达模式的高分辨率时空图谱和相应基因产物的亚细胞定位至关重要。报告基因（如GFP）与染色体背景下感兴趣的基因之间的翻译融合提供了该基因表达模式的最准确信息。在拟南芥缺乏高效的同源重组系统的情况下，利用大肠杆菌同源重组，可以精确地用荧光蛋白标记拟南芥人工大染色体，然后转移到拟南芥基因组中。本项目的主要目的是检验该细菌重组系统在拟南芥中大规模基因功能分析的效率和有效性。获得的信息的准确性，因此，该系统的效用将进行评估，生成gfp标记的拟南芥系，用于具有良好表征的表达模式的基因，以及具有明确定义的突变表型。过程中每一步的效率和“可扩展性”将被检验到全基因组水平。最后，将研究使用其他报告基因标签（如基于GAL4的报告基因标签）的效用，探索使用生成的标记克隆赋予任何感兴趣的基因特定的定义良好的表达模式的潜力，并测试特定表达模式的功能相关性。最后，将研究该技术在其他植物物种中的可移植性。更广泛的影响：在这个项目中产生的所有生物材料，经过测试后，将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、标签位置和项目进展将在项目专用网页上公布。重要的是，该项目的长期潜在影响是生产生成拟南芥“定位组”的必要工具，即所有拟南芥基因的高分辨率表达图谱。该项目还将为吸引和向高中生和本科生介绍功能基因组学领域提供一个极好的机会，特别是那些来自代表性不足的群体，特别是西班牙裔的学生。

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.