A RESOURCE CENTER FOR TETRAHYMENA THERMOPHILA

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。描述(申请人提供)：这项提议寻求建立一个基因不同的嗜热四膜虫菌株的中央储存库，四膜虫是一种有纤毛的原生动物，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，向广大社区提供关于这些菌株的基本信息；以及3)向世界各地的研究人员提供活跃生长的培养物。这一资源将为过去几十年开发的大约4000个独特的菌株提供一个安全的避风港，同时也是正在进行的四膜虫基因组测序项目的重要补充。这种由美国国立卫生研究院资助的大规模测序工作产生的活动预计将导致许多新菌株的开发，这些菌株将从中央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，该提案还打算为创建四膜虫基因组的物理图谱奠定基础。第二个目标是首席调查员实验室目前正在进行的研究的扩展，并试图使Cre-lox重组酶技术适用于四膜虫系统，以创建跨越微核基因组的中等大小(~4-5Mb)重叠缺失的小组。这一组有针对性的中型缺失将扩大我们实验室通过随机突变构建的现有的大小缺失小组，并将作为遗传和分子标记可以快速比对的支架。只有由于四膜虫独特的二态核结构，才有可能创造出几乎任何大小的可行的纯合子缺失，这是一个强大的工具，将为该系统中的功能基因组研究铺平道路，并提高资源的整体利用率。虽然四膜虫是一种生长要求简单的单细胞生物体，但它具有独特的遗传特性，以及与高等后生动物相当的结构和功能复杂性。不足为奇的是，它极大地促进了我们对广泛生物现象的理解，包括许多与人类健康直接相关的现象。这项提议中所述的资源将确保继续利用这一有机体造福整个社会。

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.