Acquisition of Equipment for High-throughput Genomics Studies

在过去的几年里，由于技术的突破，对整个基因组的分析已经成为可能，使得基因分离、鉴定和功能分析变得比以前更快和更有效。“基因组学”被广泛地定义为研究基因组的结构，这些基因组中存在的基因及其表达，以及由这些基因编码的蛋白质的功能。全基因组基因表达和功能研究的关键是使用最新开发的机器人方法来高通量地操纵基因和分析序列。亚利桑那大学的菌落和空斑采集设备(机器人工作站)、DNA序列扩增设备(聚合酶链式反应仪器)、高通量DNA测序设备(毛细管DNA测序仪)和DNA微阵列分析设备(微阵列扫描仪)将用于高通量基因组分析。菌落和菌斑拾取机器人将用于从针对不同真核和原核生物和组织的cDNA、EST和基因组文库中高通量分离单个菌落和菌斑。CDNA/EST文库将在各种条件下产生，以诱导不同的表达模式，从而识别新的基因。该扩增仪器将用于高通量生产适用于DNA测序和DNA微阵列的扩增DNA分子。毛细管测序仪将用于从各种文库插入物中扩增的DNA分子的测序。该阵列扫描仪将用于利用真核细胞和组织在不同环境条件下制备的荧光“靶”分子进行杂交后的微阵列分析。仪器对研究领域的影响在植物科学领域将特别深远。美国国家科学基金会资助的四大植物基因组基金为亚利桑那大学提供研究资金。鉴于这笔资金提供了高通量的仪器设备，预计这些项目的进展会有相当大的增长。具体地说，该仪器将允许生产和测序比这些建议最初设想的更多的cDNA库和EST文库。这应该会导致发现更多针对不同生物体的基因，并更深刻地理解基因表达是如何调控的。该仪器还将缓解基于聚合酶链式反应的微阵列生产和微阵列分析的瓶颈，这反过来将促进对通过功能基因组学协调基因表达的理解。这些研究和培训小组代表了亚利桑那大学已经参与基因组分析的小组。其他联合国组织准备开始这样的工作，作为他们正在进行的联邦资助活动的合乎逻辑的延伸。建议中的设备的提供将直接帮助他们的计划，并增加考虑采用高通量基因组学方法来解决他们特定研究问题的教职员工群体的数量。在教育和外展方面，美国大学有明确的使命，教育亚利桑那州和全国未来的劳动力。拟议设备的供应将提供宝贵的教学和培训资源。

Abstract Galbraith During the last few years, the analysis of entire genomes has been made possible by technological breakthroughs, such that gene isolation and characterization and functional analyses have become dramatically faster and more efficient than previously possible. "Genomics" is broadly defined as research toward understanding the structure of genomes, the genes present in these genomes and their expression, and the functions of the proteins encoded by these genes. Essential to genome-wide gene expression and function studies is the use of recently developed robotic methods for the high-throughput manipulation of genes and the analysis of sequences. Equipment for colony and plaque picking (a robotic workstation), for DNA sequence amplification (PCR instruments), for high-throughput DNA sequencing (a capillary DNA sequencer), and for analysis of DNA microarrays (a microarray scanner) will be used for high-throughput genomics analysis at the University of Arizona. The colony and plaque-picking robot will be used for the high-throughput isolation of individual colonies and plaques from cDNA, EST, and genomic libraries produced for different eukaryotic and prokaryotic organisms and tissues. cDNA/EST libraries will be produced under a variety of conditions designed to induce different expression patterns, thereby identifying new genes. The PCR amplification instruments will be employed for the high-throughput production of amplified DNA molecules suitable for DNA sequencing and DNA microarraying. The capillary sequencer will be employed for the sequencing of the DNA molecules that have been amplified from the various library inserts. The array scanner will be employed for the analysis of microarrays following hybridization using fluorescent "target" molecules prepared from eukaryotic cells and tissues subjected to different environmental conditions. The impact of the instrumentation on the field of research will be particularly profound in the area of the Plant Sciences. Four major NSF-funded Plant Genomics grants provide research funding for the University of Arizona. Given the availability of high-throughput instrumentation provided by this funding, a considerable increase in the progress of these projects is anticipated. Specifically, the instrumentation will permit the production and sequencing of many more cDNA and EST libraries than originally envisaged by these proposals. This should lead to greater numbers of discovered genes for different organisms, as well as a more profound understanding of how gene expression is regulated. The instrumentation will also alleviate bottlenecks in PCR-based microarray production and microarray analysis, and this in turn will accelerate understanding of the coordination of gene expression through functional genomics. These research and training groups represent ones at the University of Arizona that are already involved in genomics analyses. Other U.A. groups are poised to start such work as a logical extension of their ongoing, federally funded activities. Availability of the proposed equipment will directly assist their programs, and also increase the numbers of faculty groups considering a high-throughput genomics approach to their specific research problems. In terms of education and outreach, the U.A. has a defined mission in educating the future work force of Arizona and the Nation. The availability of the proposed equipment will provide an invaluable teaching and training resource.