Acquisition of Instrumentation for Automated DNA Analyses

我们建议建立一个DNA分析设施，购买几台用于DNA分子自动分析的关键仪器。所要求的仪器是商业上可以买到的，代表了其他主要研究型大学许多基因组和DNA测序中心的核心。这种类型的设备对于需要在全基因组范围内进行实验的研究项目来说是必不可少的。俄勒冈大学目前有三个这样的研究项目；所有这些项目都绝对需要建立拟议的DNA分析设施，才能实现其声明的目标。此外，收购所要求的仪器设备可能会产生重大的辅助效益。通过将自动化方法整合到他们的研究计划中，许多其他仪器的用户将能够更高效、更经济地完成他们的项目。DNA分析设施的建立也将极大地帮助我们招聘对发育遗传学、分子神经科学和分子进化有研究兴趣的新教师。这些都是我们计划在不久的将来招聘新教师的领域。具体来说，需要资金用于：1)集成分离系统Autogen 740，它可以在24小时内准备多达200个病毒DNA、细菌质粒DNA、酵母人工染色体(YAC)DNA或基因组DNA样本；2)Biomek 2000机器人工作站和侧载机，用于机器人操作克隆和核酸；以及3)ABI 377 DNA测序仪和必要的附件，以进行高通量自动DNA序列和微卫星分析。此外，还需要几件小型设备(包括Macintosh PowerPC 8100、两台GeneAmp 9600热循环仪和两台微型熔断器)来装备DNA分析设施。目前，俄勒冈州没有一家设施可以为拟议的DNA分析设施提供研究支持。资助这项建议将使教员能够充分开发最近启动的项目，这些项目利用复杂(数量)性状的遗传分析方面新出现的智力和技术进步，并利用基于图谱的克隆策略来分离和表征主要效应的基因。这些项目都旨在阐明基本生物学过程的遗传和分子基础，涵盖从进化和种群遗传学到发育遗传学再到行为遗传学的广泛生物学领域。例如，将由这些仪器支持的植物遗传学项目将识别植物基因组中的基因，这些基因控制着单个物种内不同植物繁殖策略的进化，在该物种中，个体在地理上被隔离了许多代。狗项目使用了与植物项目相同的实验逻辑，但在这种情况下，计划是利用狗品种之间惊人的形态和行为差异来洞察这些差异的遗传和分子基础。斑马鱼的研究工作集中在识别对生物体胚胎发育至关重要的基因，包括神经系统正常发育和功能所需的基因。这些研究人员正在寻求的一种重要方法是识别影响所述发育过程的新突变。将这些新突变放到高质量的基因图谱上的能力将有助于克隆这些基因，并最终从分子上了解这些基因是如何发挥作用的。应该指出的是，本提案中描述的许多项目都是协作性的。这反映了我们的教员努力结合我们的个人专业知识，以便更快地推进我们感兴趣的领域。通过获得所需的仪器来建立DNA分析设施将代表着这些合作努力的重大进步，并将为我们的研究计划的成功提供必要的基础设施。

We propose to establish a DNA analysis facility with the purchase of several key instruments for the automated analysis of DNA molecules. The instrumentation requested is commercially available and represents the core of many genome and DNA sequencing centers at other major research universities. This type of facility is essential for research projects that require experiments to be done on a genome-wide scale. There are currently three such research programs at the University of Oregon; all of these absolutely require the establishment of the proposed DNA analysis facility if they are to achieve their stated goals. In addition, the acquisition of the requested instrumentation is likely to have significant ancillary benefits. Many other users of the instruments will be able to complete their projects more efficiently and economically by incorporating the automated methods into their research programs. The establishment of the DNA analysis facility should also greatly aid our recruitment of new faculty with research interests in developmental genetics, molecular neuroscience and molecular evolution. These are all areas in which we plan to hire new faculty in the near future. Specifically, funds are requested for: 1) an Integrated Separation Systems AutoGen 740 that can prepare up to 200 DNA samples of viral DNA, bacterial plasmid DNA, yeast artificial chromosomal (YAC) DNA or genomic DNA in a 24 hour period; 2) a Biomek 2000 robotic workstation and sideloader for robotic manipulations of clones and nucleic acids; and 3) an ABI 377 DNA sequencer and the necessary accessories to perform high throughput automated DNA sequence and microsatellite analysis. In addition, several small equipment items (including Macintosh PowerPC 8100, two GeneAmp 9600 thermocyclers, and two microfuges) are requested to equip the DNA analysis facility. Presently, there is no facility in the State of Oregon that can provide the research support of the proposed DNA analysis facility. Fundin g this proposal will allow the faculty to fully develop recently initiated projects that utilize emerging intellectual and technical advances in the genetic analysis of complex (quantitative) characters, and that take advantage of map-based cloning strategies to isolate and characterize genes of major effect. These projects all aim to elucidate the genetic and molecular basis of fundamental biological processes, and encompass a broad spectrum of biology, from evolution and population genetics, to developmental genetics, to behavioral genetics. For example, the plant genetics project that will supported by these instruments will identify genes within the plant genome that control the evolution of different strategies of plant reproduction within a single species in which individuals have been geographically isolated for many generations. The dog project uses the same experimental logic being applied in the plant project, but in this case, the plan is to take advantage of the striking morphological and behavioral differences between dog breeds to gain insight into the genetic and molecular basis for these differences. The zebrafish research efforts are concentrated on identifying genes important for embryonic development of the organism, including genes required for the proper development and functioning of the nervous system. An important approach which is being pursued by these investigators is to identify new mutations that effect developmental processes in question. The ability to place these new mutations onto a high quality genetic map will facilitate the cloning of the genes and eventual molecular understanding of how these genes function. It should be noted that many of the projects described in this proposal are collaborative in nature. This reflects the efforts of our faculty to combine our individual expertise in order to advance our fields of interest more rapidly. Establishing the DNA analysis facility with the acquisition of the requested instrumentati on will represent a major advance in these collaborative efforts, and will provide the infrastructure necessary for the success of our research programs.