A RESOURCE CENTER FOR TETRAHYMENA THERMOPHILA

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。描述（由申请人提供）：本提案旨在为嗜热四膜虫（Tetrahymena thermophila）建立一个遗传上不同的菌株的集中存储库，四膜虫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。库存中心将1)收集、注释和存储实验有用的细胞系；2)建立一个数据库，向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来开发的大约4000种独特菌株提供安全的避风港，同时，作为正在进行的四膜虫基因组测序计划的重要辅助。这项由美国国立卫生研究院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的物理图谱奠定基础。第二个目标是目前在首席研究员实验室进行的研究的延伸，并寻求将Cre-lox重组酶技术应用于四膜虫系统，以便在微核基因组中创建一个中等大小（~ 4-5 Mb）重叠缺失的面板。这组特异性靶向、中等大小的缺失将增强我们实验室已经通过随机诱变构建的大大小小的缺失面板，并将作为一个支架，在其上可以快速排列遗传和分子标记。由于四膜虫独特的二态核结构，创造几乎任何大小的可行纯合缺失的能力是唯一可能的，并且代表了一个强大的工具，将为该系统的功能基因组研究铺平道路，并提高资源的整体利用率。虽然是单细胞生物，生长要求简单，但四膜虫具有独特的遗传特性，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一系列生物现象的理解，其中包括许多与人类健康直接相关的现象。本提案中描述的资源将确保为整个社会的利益继续使用这种生物。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.