A Resource Center for Tetrahymena Thermophila

描述(由申请人提供)：本提案寻求继续为四膜虫种群中心和辅助四膜虫基因组数据库(TGD)提供资金，以维持其目前的运营并扩大其作为整个社区资源的能力。嗜热四膜虫是真核细胞和分子生物学的重要模型，它有助于我们理解一系列与人类健康和疾病直接相关的生物现象，包括癌症、不孕不育、衰老和神经内分泌功能障碍。四膜虫还显示出作为蛋白质制造平台的巨大前景，包括生产流感、疟疾和其他传染病病原体的疫苗。具体地说，我们的目标是1)在我们现有的设施中保持基因库存，并通过我们简单的基于网络的订购系统将它们提供给科学界；2)扩大现有菌株的现有收集和完整注释；3)扩展中心的服务，包括存储和传播有用的质粒构建体，以及一项新的收费转化服务，将显著增加成本回收；4)提供短期技术培训课程，以促进新系统的研究人员使用四膜虫；5)通过TGD Wiki促进继续基于社区的四膜虫大核基因组注释；6)建立库存中心的单个菌株与TGD相关基因页面之间的链接，简化有用菌株的鉴定，并改善中心用户的信息获取。最后，作为股票中心研究部分的一部分，我们将创建更有效的微核转化策略，并利用这些策略在所有五条微核染色体上产生一组小的、有针对性的缺失。这个小组将作为一个脚手架，遗传和分子标记可以在其上快速比对，并将有力地支持目前在嗜热锥虫中进行的许多功能基因组研究。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存活的纯合子缺失，并代表了这种重要模式生物的许多新特征之一。

DESCRIPTION (provided by applicant): This proposal seeks continued funding for the Tetrahymena Stock Center and ancillary Tetrahymena Genome Database (TGD) in order to maintain their current operations and expand their capabilities as resources for the community at-large. A key model for eukaryotic cell and molecular biology, Tetrahymena thermophila has been instrumental to our understanding of a wide range of biological phenomena with direct relevance to human health and disease including cancer, infertility, aging, and neuroendocrine dysfunction. Tetrahymena has also shown great promise as a platform for protein manufacture including the production of vaccines for influenza, malaria and other infectious agents. Specifically, we aim to 1) maintain genetic stocks in our current facilities and make them available to the scientific community via our simple web-based ordering system; 2) expand the current collection and complete annotation of existing strains; 3) extend the Center's services to include storage and dissemination of useful plasmid constructs, and a new fee-based transformation service that will significantly increase cost recovery; 4) offer short techniques an methodology training sessions to promote the use of Tetrahymena by investigators who are new to the system; 5) promote continued community-based annotation of the Tetrahymena macronuclear genome via the TGD Wiki; 6) establish links between individual strains in the Stock Center collection and relevant gene pages in TGD to simplify identification of useful strains and improve information access for the Center's users. Lastly, as part of the research component of the Stock Center, we will 7) create more efficient strategies for micronuclear transformation and utilize these to generate a panel of small, targeted deletions in all five micronuclear chromosomes. This panel will act as a scaffold on which genetic and molecular markers can be quickly aligned and will strongly support the many functional genomic studies now underway in T. thermophila. 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 one of many novel features of this important model organism.