DEVELOPMENT OF A PRIMATE GENOMIC RESOURCE

这项修订后的R24资源相关研究资助的竞争性续订侧重于创建 可应用于非人类灵长类动物研究的蛋白质组资源。这项建议是建立在 在最初的资助期间构建的资源，在此期间我们培育了广泛的恒河猴 (猕猴)特有的基因组工具。通过构建互补的蛋白质组资源， 我们打算向研究界提供物种特有的全谱基因组和 将功能基因组学技术应用于非人类所需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研究所合作 太平洋西北国家实验室理查德·D·史密斯博士领导的光谱分析小组。这群人 为这项工作带来无与伦比的质谱学仪器和尖端的蛋白质组学专业知识。在……里面 目标1，我们将使用鸟枪蛋白质组学和傅里叶变换离子回旋共振质谱学来 建立猕猴特异性蛋白质数据库，用于鉴定各种猕猴体内存在的蛋白质 纸巾。这些数据库将用于AIM 2，以表征发生在AIM 2中的蛋白质谱变化 对猴免疫缺陷病毒(SIV)感染的反应。我们将从分析外周血开始 体外感染SIVmac239的单个核细胞(PBMC)。然后我们将感染相同的动物从 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STRATE 同位素标记及结合AMT的无标记相对绝对定量方法 标签法。这些研究将有助于推动蛋白质组资源的开发，并增强我们的 了解使用猕猴作为艾滋病研究的模型。在目标3中，我们将制定 蛋白质组数据的存储、解释和传播所需的生物信息学基础设施， 整合了许多开源工具，包括LabKey。该信息资源将提供Web访问 在整个过程中，结果层次分明。作为我们努力将蛋白质组学的使用扩展到 非人灵长类研究社区，我们还将主办年度研讨会，研究人员可以讨论 如何利用蛋白质组数据库，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.