Chromatin Charting: Organization and Dynamics of Plant Nuclear DNA in situ

染色质图谱：植物核 DNA 的原位组织和动态

• 批准号: 9872636
• 负责人: Eric Lam
• 金额: $ 67.62万
• 依托单位: Rutgers University New Brunswick
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-10-01 至 2000-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9872636&HistoricalAwards=false
• 关键词: Chromatin; Charting; Organization; Dynamics; Plant

The nucleus is the subcellular organelle in which the bulk of the genomic information within an eukaryotic cell is organized. Although tremendous advances in sequencing genomes of organisms have taken place in the past few years, the way by which this vast amount of genetic information is stored and physically manipulated in the nuclei of plants and animals remains a mystery. From studies using hybridization technologies and microscopy work, a picture of an organized subnuclear structure has emerged. One major subnuclear structure is the chromosomes, which contain the genome of an organism. More recently, the application of the Green Fluorescent Protein (GFP) as an in vivo tag of chromosomal substances has allowed the visualization of chromosomes in live cells of animals and fungi. The goal for this project is to extend the GFP technology to understand subnuclear architecture by charting the relative physical position of specific genes on each of the chromosomes using cells within an intact plant. To achieve the objective of visualizing and charting the genes on all the chromosomes of a model plant, Arabidopsis, GFP and two different color variants (yellow [YFP] and blue [BFP]) of this protein will be used as in vivo tags in transgenic plants. Using the three distinct markers, the physical organization of the genome in relation to gene expression and gene mobilization patterns will be characterized. The project is a first step to establish this technology as a powerful tool to visualize the relationship between the genome organization and gene expression. While it is being tested in a model plant, Arabidopsis, this approach should provide the necessary resources and tools to address fundamental questions about subnuclear architecture and chromatin dynamics in all plants. Further, insights from this work should contribute to our understanding of global gene regulation as well as epigenetic phenomena in all eukaryotic organisms.

细胞核是亚细胞细胞器，真核细胞内的大量基因组信息在其中组织。尽管过去几年在生物体基因组测序方面取得了巨大进展，但动植物细胞核中大量遗传信息的存储和物理操作方式仍然是个谜。通过使用杂交技术和显微镜工作的研究，一幅有组织的亚核结构的图片已经出现。染色体是一种主要的亚核结构，它包含生物体的基因组。 最近，绿色荧光蛋白（GFP）作为染色体物质的体内标签的应用使得动物和真菌的活细胞中的染色体得以可视化。该项目的目标是通过使用完整植物内的细胞绘制每条染色体上特定基因的相对物理位置图来扩展 GFP 技术以了解亚核结构。为了实现对模型植物、拟南芥所有染色体上的基因进行可视化和绘制图表的目标，该蛋白的 GFP 和两种不同颜色的变体（黄色 [YFP] 和蓝色 [BFP]）将被用作转基因植物的体内标签。使用三个不同的标记，将表征与基因表达和基因动员模式相关的基因组的物理组织。 该项目是将这项技术打造成可视化基因组组织和基因表达之间关系的强大工具的第一步。 虽然它正在模型植物拟南芥中进行测试，但这种方法应该提供必要的资源和工具来解决所有植物中亚核结构和染色质动力学的基本问题。此外，这项工作的见解应有助于我们理解所有真核生物的全球基因调控以及表观遗传现象。