A Resource Center for Tetrahymena Thermophila

这项建议旨在建立一个集中的储存库，储存遗传上不同的菌株， Tetmhymena thermophila，一种纤毛原生动物，已成为真核生物研究的关键模型。 细胞和分子生物学超过50年。库存中心将1）收集、注释和存储 实验上有用的细胞系; 2）建立一个数据库，将提供有关这些的基本信息 菌株的社区在广大;和，3）供应积极增长的文化，以研究人员在世界各地。 这一资源将为过去几年中开发的大约4000种独特菌株提供安全的避风港。 几十年，同时，作为一个重要的附属正在进行的四膜虫基因组 测序项目。这项由NIH资助的大规模测序工作所产生的活动预计将 导致许多新菌株的发展，这将大大受益于集中的存在， 储存和分配设施。除了建立股票中心外，该提案还打算 为绘制四膜虫基因组物理图谱奠定了基础。第二个目标是 主要研究者实验室目前正在进行的研究的扩展，并寻求 使Cre-lox重组酶技术适应四膜虫系统，以创建一组中等大小的 （~ 4-5 Mb）重叠缺失跨越微核基因组。这是一个有针对性的，中... 大小的删除将增加现有的面板的大小删除已经构建在我们的 实验室通过随机诱变，并将作为一个支架上的遗传和分子标记，可以 快速对齐。创建几乎任何大小的可行的纯合缺失的能力仅是可能的， 四膜虫独特的二形核结构，并代表了一个强大的工具，将铺平道路， 为功能基因组研究提供了一种新的途径，提高了资源的整体利用率。虽然 四膜虫是一种生长条件简单的单细胞生物， 属性，以及结构和功能的复杂性，可与高等后生动物相媲美。不 令人惊讶的是，它极大地促进了我们对各种生物现象的理解 包括许多与人类健康直接相关的问题。本提案中所述的资源将确保 为了整个社会的利益而继续使用这种生物。

This proposal seeks to establish a centralized repository for genetically distinct strains of Tetmhymena thermophila, a ciliated protozoan that has served as a key model for studies of eukaryotic cellular and molecular biology for more than 50 years. The stock center will 1) collect, annotate, and store experimentally useful cell lines; 2) establish a database that will provide essential information about these strains to the community at-large; and, 3) supply actively growing cultures to researchers around the world. This resource will provide a safe haven for the roughly 4000 unique strains developed over the past several decades, and at the same time, serve as an important adjunct to the ongoing Tetrahymena genome sequencing project. Activities spawned by this large-scale, NIH-funded sequencing effort are expected to result in the development of many new strains that will benefit greatly from the existence of a centralized storage and distribution facility. In addition to the establishment of a stock center, the proposal also intends to lay the groundwork for creation of a physical map of the Tetrahymena genome. This second goal is an extension of research currently being undertaken in the Principal Investigator's laboratory, and seeks to adapt the Cre-lox recombinase technology to the Tetrahymena system in order to create a panel of mid-sized (~ 4-5 Mb) overlapping deletions spanning the micronuclear genome. This set of specifically targeted, mid- sized deletions will augment an existing panel of large and small deletions already constructed in our laboratory by random mutagenesis, and will act as a scaffold on which genetic and molecular markers can be quickly aligned. The ability to create viable, homozygous deletions of virtually any size is only possible due to the unique dimorphic nuclear structure of Tetrahymena, and represents a powerful tool that will pave the way for functional genomic studies in this system and enhance the utility of the resource overall. Although a unicellular organism with simple growth requirements, Tetrahymena is endowed with unique genetic properties, and a structural and functional complexity comparable to that of higher metazoa. Not surprisingly, it has contributed enormously to our understanding of a wide range of biological phenomena including many with direct relevance to human health. The resource described in this proposal will ensure the continued use this organism for the benefit of society as a whole.