Global Expression Studies of the Arabidopsis Genome

到2000年底，对130Mb拟南芥基因组DNA序列的阐明估计揭示了使用各种计算方法的大约25000个基因的存在。为了让这些庞大的结构信息对植物生物学家最有用，还需要另一个大规模的项目来表征整个基因组的表达模式。该项目将为鉴定所有转录单位和构建拟南芥基因组中所有基因的全长cDNA提供一个框架。这样的努力将使精确确定每个转录单位编码的每种蛋白质的分子质量成为可能。此外，这种方法比让不同的实验室分离和测序感兴趣的单个cDNA更具成本效益。该项目的中心焦点是通过利用新兴的基因组技术来了解植物生长和发育的各个方面，为整个植物生物界开发分子资源(基础设施)。该项目利用拟南芥基因组序列的快速增长和新技术的出现来实现这一长期目标。该奖项将支持以下目标：(1)构建和开发包含拟南芥基因组倡议所有预测基因的高密度Affymetrix寡核苷酸芯片。该芯片将每年更新，直到整个基因组序列可用。(2)利用Affymetrix芯片和从暗植物和光植物不同处理条件下分离的约100个不同组织中分离的mRNAs，进行拟南芥所有基因表达模式的全局表达研究。这些芯片将提供给整个科学界(3)对8000个全长DNA进行分离和测序。这一目标的完成将提供1/3的拟南芥基因的实验性注释，并确定目前用于基因预测的计算方法的准确性。这项提议的一个重要特点是，所有上述材料一旦生产出来，就将免费提供给植物生物界。这些新资源将促进植物生物学的发展，进而推动世界农业和植物生物技术的发展。

The elucidation of the DNA sequence of the 130 Mb Arabidopsis genome by the end of the year 2000 is estimated to reveal the presence of approximately 25,000 genes using various computational approaches. In order for this vast structural information to be most useful to plant biologists, another large-scale project is needed to characterize the expression patterns of the entire genome. This project will provide a framework for the identification of all the transcriptional units and the construction of full-length cDNAs for all the genes in the Arabidopsis genome. Such an endeavor will allow the precise determination of the molecular mass of each protein encoded by each transcriptional unit. In addition, this approach is much more cost effective than having different laboratories isolate and sequence individual cDNAs of interest. The central focus of this project is the development of molecular resources (infrastructure) for the entire plant biology community by utilizing emerging genomic technologies for understanding various aspects of plant growth and development. The project takes advantage of the rapid growth in Arabidopsis genomic sequence and the advent of new technologies to achieve this long term goal.This award will support the following objectives: (1) Construction and development of a high density Affymetrix oligonucleotide chip that contains all the predicted genes from the Arabidopsis Genome Initiative. The chip will be updated annually until the entire genome sequence is available. (2) Global expression studies for determining the expression pattern of all the Arabidopsis genes utilizing the Affymetrix chip and mRNAs isolated from ~100 different treatment conditions of various tissues from dark and light grown plants. The chips will be available to the entire scientific community (3) Isolation and sequencing of 8,000 full-length cDNAs. The completion of this goal will provide experimental annotation of 1/3 of all Arabidopsis genes and determine the accuracy of computational approaches currently utilized for gene prediction. The cDNAs will be useful tools for a variety of experimental purposes.An important feature of this proposal is that all of the above-described materials will be made freely available to the plant biology community as soon as they are produced. These new resources will allow the advancement of plant biology and by extension, world agriculture and plant biotechnology.