SGER: Global Analysis of the Nuclear Genome

该项目的目标是找出植物细胞核的总蛋白质组成，并观察这种组成是否在不同的植物器官和细胞类型中发生变化。为了实现这一目标，PI将联合收割机结合他们在细胞核高速荧光激活流式分选和高通量分析蛋白质组学领域的技术专业知识。该实验将使用流式分选技术从植物组织和器官制备的匀浆中快速纯化细胞核。这些细胞核将使用高通量蛋白质组学技术，特别是质谱法进行分析，以识别细胞核内包含的尽可能多的单个蛋白质。该项目将使用两种植物物种开发这些程序，双子叶模式拟南芥和单子叶作物水稻。选择这些物种是因为它们的完整核基因组序列是可用的。进一步的实验将检查特定器官（根、芽、叶、花）和根的特定细胞（表皮、内皮层、维管系统、分生组织和皮层）内的核蛋白组成。SGER资金将用于确定该实验策略的技术可行性。

The goal of the project is to find out the total protein composition of the plant nucleus, and to see whether this composition changes in different plant organs and cell types. To reach this goal, the PIs will combine their technical expertise in the areas of high-speed fluorescence-activated flow sorting of nuclei and high throughput analytical proteomics. The experiments will use flow sorting to rapidly purify nuclei from homogenates prepared from plant tissues and organs. These nuclei will be analyzed using high throughput proteomic techniques, particularly mass spectrometry, to identify as many as possible of the individual proteins contained within the nuclei. The project will develop these procedures using two plant species, the dicot model Arabidopsis thaliana, and the monocot crop Oryza sativa. These species were chosen since their complete nuclear genome sequences are available. Further experiments will examine nuclear protein composition within specific organs (roots, shoots, leaves, flowers), and within specific cells of the root (epidermis, endodermis, vascular system, meristem, and cortex). The SGER funding will be employed to determine the technical feasibility of this experimental strategy.