A Resource Center for Tetrahymena Thermophila

描述（由申请人提供）： 这项建议旨在建立一个集中的储存库，用于储存嗜热四膜虫的遗传上不同的菌株，这是一种纤毛原生动物，50多年来一直是真核细胞和分子生物学研究的关键模型。储备中心将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，为整个社区提供有关这些菌株的基本信息; 3）为世界各地的研究人员提供积极生长的培养物。这一资源将为过去几十年中开发的大约4000种独特菌株提供安全的避风港，同时也是正在进行的四膜虫基因组测序项目的重要辅助手段。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开发，这些菌株将从集中存储和分配设施的存在中受益匪浅。除了建立一个库存中心外，该提案还打算为创建四膜虫基因组物理图谱奠定基础。这第二个目标是目前在主要研究者的实验室进行的研究的延伸，并寻求使Cre-lox重组酶技术适应四膜虫系统，以创建一组跨越微核基因组的中等大小（~ 4-5 Mb）重叠缺失。这组特异性靶向的中等大小的缺失将增加我们实验室中已经通过随机诱变构建的现有大小缺失组，并将作为遗传和分子标记可以快速对齐的支架。创建几乎任何大小的可行的纯合缺失的能力是唯一可能的，由于四膜虫独特的二态核结构，并代表了一个强大的工具，将铺平道路，在这个系统中的功能基因组研究，并提高资源的整体效用。虽然四膜虫是一种生长条件简单的单细胞生物，但它具有独特的遗传特性，其结构和功能的复杂性可与高等后生动物相媲美。毫不奇怪，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生物现象。本提案中描述的资源将确保继续使用这种生物体，以造福整个社会。

DESCRIPTION (provided by applicant): This proposal seeks to establish a centralized repository for genetically distinct strains of Tetrahymena 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, midsized 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.