An Automated DNA Sequencer for Analysis of Gene Structure and Function

此应用程序请求对自动DNA开发的部分支持 测序设施。 华盛顿大学生物化学系、植物学系、遗传学系和动物学系的七个独立实验室（主要用户）和这些系的另外七个实验室（次要用户）正在参与这项提案，对这些实验室而言，DNA测序是一个主要的研究重点，但对DNA测序的需求不太广泛。这一应用的动力来自于这些人的认识，即现有的DNA手动测序方法比采用荧光化合物跟踪DNA片段的自动化方法更慢，工作更密集，更昂贵。参与这项申请的所有14名教职员工都得出结论，需要依赖手动方法进行DNA测序，这对他们的研究进展产生了明显的负面影响。 构成该申请的14个项目都由国家科学基金会，国立卫生研究院和其他授予机构的一项或多项研究赠款。主要项目包括：果蝇卵子发生过程中细胞迁移所需基因的分析（C。贝格）;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）;和昆虫发育的激素调节（L. Riddiford）。小项目包括：拟南芥revolta基因的特征。COMAI）; S.酿酒酵母（T.戴维斯）;酵母细胞周期的遗传控制（L。Hartwell）;调节 非洲爪蟾和斑马鱼的中胚层图案化（D. Kimelman）;果蝇发育中极性的起源（H. Ruohola-Baker）;果蝇异染色质基因和位置效应（B. akimoto）;以及酵母中调节蛋白及其靶基因的进化（E.Young）。 我们请求购买377型ABI Prism自动DNA测序仪及相关计算机软件，以及Genetics PCR System 9600。 这些仪器将存放在生物化学系提供的空间内。该设施将在按服务收费的基础上运行（成本中心）。预计参与实验室将利用该设施80-90%的能力，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.