An Automated DNA Sequencer for Analysis of Gene Structure and Function

该应用程序请求为开发自动DNA测序设施提供部分支持。华盛顿大学生物化学、植物学、遗传学和动物学系的七个独立实验室，其中DNA测序是主要的研究重点(主要用户)，以及这些系中另外七个对DNA测序有重要但不那么广泛的需求的实验室(次要用户)，正在参与这项提案。这一应用的动力来自于这些人意识到，现有的DNA手动测序方法比使用荧光化合物跟踪DNA片段的自动化方法速度更慢、工作更密集、成本更高。所有参与这项申请的14名教职员工都得出结论，依赖手工方法进行DNA测序的需要正在对他们的研究进展产生明显的负面影响。组成这项申请的14个项目都是由国家科学基金会、国家卫生研究院和其他资助机构提供的一项或多项研究资助。主要项目包括：分析果蝇卵子发生过程中细胞迁移所需的基因(C.Berg)；哺乳动物Metaxin基因家族的结构和功能(P.Bornstein)；海洋植物系统的分子分析(R.cattolico)；鸟类MHC II类区域的大规模测序(S.Edwards)；来自不同分类群的核RNA聚合酶II的两个最大亚基的比较序列分析(B.Hall)；肌肉肌酸激酶基因的调控分析(S.Hauschka)；以及昆虫发育的激素调节(L.Riddiford)。较小的项目包括：拟南芥(L.Comai)反叛基因的特征；酿酒酵母(T.Davis)微管组织中心组件的鉴定；酵母细胞周期的遗传控制(L.Hartwell)；非洲爪哇和斑马鱼中中胚层图案的调控(D.Kimelman)；果蝇发育中极性的起源(H.Ruohola-Baker)；果蝇中异色基因和位置效应(B.Akimoto)；以及酵母中调控蛋白及其目标基因的进化(E.Young)。我们要求购买377型ABI Prism自动DNA测序仪和相关的计算机软件，以及GeneAmp PCR系统9600。这些仪器将被安置在生物化学系提供的空间中。该设施将以按服务收费的方式运行(成本中心)。预计该设施80%-90%的能力将由参与实验室使用，其余能力将以先到先得的方式提供给华盛顿大学的研究人员。

This application requests partial support for the development of an Automated DNA Sequencing Facility. Seven independent laboratories in the Departments of Biochemistry, Botany, Genetics, and Zoology at the University of Washington, for whom DNA sequencing represents a major research focus (major users), and an additional seven laboratories in these Departments with a significant but less extensive need for DNA sequencing (minor users), are participating in this proposal. The impetus for this application derives from the realization by these individuals that existing methods for manual sequencing of DNA are slower, more work-intensive, and more expensive than are automated methods that employ fluorescent compounds to track DNA fragments. All 14 faculty members involved in this application have concluded that the need to rely on manual methods for DNA sequencing is having a distinctly negative impact on the progress of their research. The 14 projects that comprise this application are all funded by one or more research grants from the National Science Foundation, National Institutes of Health, and other granting agencies. The major projects involve: an analysis of genes required for cell migrations during oogenesis in Drosophila (C. Berg); the structure and function of the mammalian metaxin gene family (P. Bornstein); a molecular analysis of marine plant systems (R. Cattolico); large-scale sequencing of the MHC class II region in birds (S. Edwards); comparative sequence analysis of the two largest subunits of nuclear RNA polymerase II from different taxa (B. Hall); regulatory analyses of muscle creatine kinase genes (S. Hauschka); and hormonal regulation of insect development (L. Riddiford). The minor projects involve: characterization of the revolta gene of Arabidopsis thaliana (L. Comai); identification of components of the microtubule organizing center in S. cerevisiae (T. Davis); genetic control of the yeast cell cycle (L. Hartwell); regulation of mesodermal patterning in Xenopus and zebrafish (D. Kimelman); origin of polarity in Drosophila development (H. Ruohola-Baker); heterochromatic genes and position effects in Drosophila (B. akimoto); and evolution of a regulatory protein and its target gene in yeast (E.Young). We request the purchase of a model 377 ABI Prism Automated DNA Sequencer with associated computer software, and a GeneAmp PCR System 9600. These instruments will be housed in space provided by the Department of Biochemistry. The facility will be run on a fee-for-service basis (cost center). It is anticipated that 80-90% of the capacity of the facility will be utilized by the participating laboratories and that the remaining capacity will be made available to investigators at the University of Washington on a first-come, first-served basis.