SGER: Developing Methods for Global Expression Analysis at Cellular Resolution

0127676优点确定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开发，以快速确定数千个转录本的细胞类型特异性表达。这项技术包括通过原生质体在特定细胞类型中表达GFP的转基因株来分离植物细胞，使用荧光激活细胞分选器(FACS)获得丰富的GFP表达细胞群，以及将从分选细胞中分离出的mRNA应用于微阵列。结果是来自特定细胞类型的表达配置文件。已经进行了广泛的控制，并完成了对单个转基因株系的整个方案的第一轮。这些结果与特定基因的微阵列图谱非常有希望，这些基因与它们在拟南芥根中独立确认的表达模式一致。这项建议的目的是：1)进一步测试和发展方法学。2)扩大已分类细胞系的数量。3)开发分析和传播数据的计算工具。与该项目相关的风险与给工厂社区带来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.