Characterizing Sub-Cellular Compartments in Maize Using Fluorescent Protein Tagged Lines

玉米细胞由相互连接但离散的室室组成，这些室室有助于维持细胞功能和秩序。鉴定定位于这些区室的蛋白质对于理解玉米的发育和生理过程至关重要，从而为作物改良提供指导性信息。该项目利用基因组学的最新进展来鉴定定位于不同细胞区室的蛋白质。该项目将生成表达带有荧光标记的蛋白质的报告细胞系。使用共聚焦显微镜，这些线条将显示有关标记蛋白质表达的时间和地点以及特定蛋白质如何相互作用和功能的视觉信息。用于标记的基因将提供完整的细胞区室标记覆盖。一些资源将用于基因选择，包括来自生长素/细胞分裂素激素途径和来自rab介导的囊泡运输途径的已知基因。基因选择也将根据玉米基因组测序项目产生的公共基因模型和玉米研究界的具体要求进行指导。该项目的结果将包括一组稳定的标记系，表达100种蛋白质的荧光蛋白衍生标签，这将代表细胞区室的完全覆盖。标记的基因结构将从杰克逊实验室免费获得，转基因品种的种子将从玉米遗传库存中心获得。获取项目成果：细胞系特征在细胞水平上的数据将汇编在一份发布在公共网站http://maize.tigr.org/cellgenomics上的本地化目录中。细胞生物学讲习班将帮助培训科学用户群体使用共聚焦显微镜分析标记细胞系。该项目还将通过加强细胞和分子生物学的教育，为落基山脉西部的部落社区学院提供服务。因此，该项目将产生桥梁细胞生物学和功能基因组学的资源，并将在不同的科学和学习社区提供培训。培训和推广活动将在植物基因组研究推广门户网站http://www.plantgdb.org/pgrop/pgrop.php上报告。

Maize cells consist of interconnected but discrete compartments that help to maintain cellular function and order. Identifying proteins that localize to these compartments is critical to understanding developmental and physiological processes in maize, which in turn provides guiding information for crop improvement. This project makes use of recent advances in genomics to identify proteins that localize to diverse cellular compartments. The project will generate reporter lines that express proteins tagged with a fluorescent marker. Using confocal microscopy, the lines will display visual information about when and where the tagged proteins are expressed and how specific proteins may be interacting and functioning. Genes for tagging will provide complete marker coverage of cellular compartments. Several resources will be used for gene selection, including known genes from the auxin/cytokinin hormone pathway and from the RAB-mediated vesicle trafficking pathways. Gene selections will also be guided by public gene models generated from maize genome sequencing projects and by specific requests from the maize research community. The outcomes of the project will include a set of stable tagged lines expressing fluorescent protein-derived tags for 100 proteins, which will represent full coverage of cellular compartments. The tagged gene constructs will be freely available from the Jackson lab, and seeds of transgenic lines will be available from the Maize Genetics Stock Center. Access to project outcomes:Data on characterization of the lines at the cellular level will be compiled in a localization catalog posted on a public website at http://maize.tigr.org/cellgenomics. Cell biology workshops will help train the scientific user community in analysis of the tagged lines using confocal microscopy. The project will also serve tribal community colleges in the Rocky Mountain West by enhancing education in cell and molecular biology. The project will therefore generate resources that bridge cell biology and functional genomics and will provide training across diverse scientific and learning communities. Training and outreach activities will be reported on the Plant Genome Research Outreach Portal at http://www.plantgdb.org/pgrop/pgrop.php.