DEVELOPMENT OF A PRIMATE GENOMIC RESOURCE

R24资源相关研究补助金的修订竞争性更新的重点是创建 蛋白质组学资源，可以应用于研究使用非人类灵长类动物。该提案建立在 在最初的资助期内，我们开发了大量的恒河猴资源， （Macaca mulatta）特异性基因组工具。通过构建互补蛋白质组资源， 我们打算为研究界提供全方位的物种特异性基因组和 将功能基因组学技术应用于非人类的蛋白质组学工具和数据库 灵长类研究为了建立这些资源，我们与生物系统分析和质量 光谱小组由理查德·D·史密斯在太平洋西北国家实验室。这群 带来了无与伦比的质谱仪器和尖端的蛋白质组学专业知识的努力。在 目的1、利用鸟枪蛋白质组学和傅立叶变换离子回旋共振质谱技术， 建立猕猴特异性蛋白质数据库，用于鉴定各种恒河猴中存在的蛋白质 组织中这些数据库将用于目标2，以表征在以下情况下发生的蛋白质谱变化： 猴免疫缺陷病毒（SIV）感染。我们将开始分析外周血 用SIVmac239体外感染的单核细胞（PBMC）。然后我们将感染相同的动物， 并分析血液、肠道相关淋巴组织和淋巴结。 感染后多个早期时间点。定量蛋白质组学分析将使用稳定的 同位素标记和无标记比较绝对定量方法与AMT的组合 标签方法。这些研究将有助于推动蛋白质组资源的开发，并提高我们的 了解使用猕猴作为艾滋病研究模型的情况。在目标3中，我们将开发 存储、解释和传播蛋白质组学数据所需的生物信息学基础设施， 包括LabKey在内的许多开源工具。此信息资源将提供Web访问， 在整个过程中的结果层次。作为我们努力将蛋白质组学的应用扩展到 非人类灵长类动物研究社区，我们还将举办年度研讨会，研究人员可以讨论 如何利用蛋白质组学数据库，并将蛋白质组学纳入自己的研究活动。

This revised competitive renewal of an R24 Resource-Related Research Grant is focused on creating proteomic resources that can be applied to research using nonhuman primates. The proposal builds upon resources constructed during the original funding period in which we developed extensive rhesus macaque (Macaca mulatta)-specific genomic tools. Through the construction of complementary proteomic resources, we intend to provide the research community with the full spectrum of species-specific genomic and proteomic tools and databases needed to apply the technologies of functional genomics to nonhuman primate research. To build these resources, we have teamed with the Biological Systems Analysis and Mass Spectrometry group headed by Dr. Richard D. Smith at Pacific Northwest National Laboratory. This group brings to the effort unrivaled mass spectrometry instrumentation and cutting-edge proteomics expertise. In Aim 1, we will use shotgun proteomics and Fourier transform ion cyclotron resonance mass spectrometry to build macaque specific protein databases for use in identifying proteins present in various rhesus macaque tissues. These databases will be used in Aim 2 to characterize the changes in protein profiles that occur in response to simian immunodeficiency virus (SIV) infection. We will begin by analyzing peripheral blood mononuclear cells (PBMCs) infected in vitro with SIVmac239. We will then infect the same animals from which the PBMCs were derived and analyze blood, gut-associated lymphoid tissue, and lymph nodes at multiple early time points after infection. Quantitative proteomic analyses will be performed using stable isotope labeling and a label-free comparative absolute quantification method in combination with the AMT tag approach. These studies will serve to drive the development of proteomic resources and to enhance our understanding of the use of macaques as models for AIDS research. In Aim 3, we will develop the bioinformatics infrastructure needed for the storage, interpretation, and dissemination of proteomics data, incorporating many open source tools including LabKey. This information resource will offer Web access to tiers of results throughout the process. As part of our efforts to extend the use of proteomics to the nonhuman primate research community, we will also host annual workshops where investigators can discuss how to make use of proteomic databases and to incorporate proteomics into their own research activities.