SGER: Developing Methods for Global Expression Analysis at Cellular Resolution

表征基因功能的关键第一步是确定基因的表达模式。一种新的方法正在植物中发展，以快速确定数千个转录本的细胞类型特异性表达。该技术包括通过在特定细胞类型中表达GFP的原生质体转基因系分离植物细胞，使用荧光激活细胞分选器（FACS）获得表达GFP的丰富细胞群，并将从分选细胞中分离的mRNA应用于微阵列。结果是来自特定细胞类型的表达谱。已经进行了广泛的控制，并且已经完成了使用单一转基因品系的第一轮整个方案。这些结果非常有希望，特定基因的微阵列图谱与它们在拟南芥根中独立确认的表达模式一致。本提案的目的是：1)进一步测试和发展该方法。2)扩大细胞系的分类数量。3)开发计算工具来分析和传播数据。与该项目相关的风险与植物群落的巨大潜在利益相平衡。

0127676BenfeyA critical first step in characterizing gene function is defining the expression pattern of a gene. A novel methodology is being developed in plants to rapidly determine the cell-type specific expression of thousands of transcripts in concert. The technique involves separating plant cells by protoplasting transgenic lines that express GFP in specific cell types, obtaining an enriched population of GFP expressing cells using a Fluorescence Activated Cell Sorter (FACS), and applying mRNA isolated from sorted cells to microarrays. The result is an expression profile from a specific cell type. Extensive controls have been performed and a first round of the entire protocol with a single transgenic line has been completed. The results are extremely promising with microarray profiles of specific genes in agreement with their independently confirmed expression patterns in the Arabidopsis root. The aims of this proposal are to: 1) Further test and develop the methodology. 2) Expand the number of sorted cell lines. 3) Develop computational tools to analyze and disseminate the data. The risk associated with this project is balanced by the enormous potential benefit to the plant community.