SGER: Developing Methods for Global Expression Analysis at Cellular Resolution

0127676 Benfey表征基因功能的关键第一步是定义基因的表达模式。一种新的方法正在植物中开发，以快速确定成千上万的转录本的细胞类型特异性表达。该技术涉及通过原生质体化在特定细胞类型中表达GFP的转基因系来分离植物细胞，使用荧光激活细胞分选仪（FACS）获得GFP表达细胞的富集群体，并将从分选的细胞分离的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，并且已经完成了具有单个转基因系的整个方案的第一轮。结果是非常有前途的微阵列配置文件的特定基因与他们的独立确认的表达模式在拟南芥根。本建议的目的是：1）进一步测试和发展方法。 2)扩大分选细胞系的数量。 3)开发分析和传播数据的计算工具。与该项目相关的风险被工厂社区的巨大潜在利益所平衡。

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.