CORE--FACS AND RESEARCH ANIMALS

这项拨款中提出的实验在很大程度上依赖于密切的合作 三个PI之间的相互作用。核心组件将极大地 促进这些互动。核心B分为3个部分：A) 转基因/基因敲除小鼠的培育，b)维持常规 转基因和基因敲除小鼠，c)流式细胞术。 核心B旨在提供老年T细胞受体转基因基因 所有三个项目所需的基因敲除和常规小鼠 这个项目。核心B将培育和筛选转基因动物(使用FAC 或聚合酶链式反应分析)，并在必要时回交以使其适合 对于该计划的个别项目中详细说明的实验。 因为这个过程是非常劳动密集型的，是一个集中化的设施 专门执行这项任务是必不可少的。核心将获得 各种老化的常规、转基因或基因敲除小鼠 个别项目，必要时予以检疫，制定标准化 转基因/敲除基因表达的筛选程序 每个病例，并将繁殖和回交的小鼠近交系。这个 目标是拥有健康的、基因同种的小鼠 该项目的调查人员。调查人员将能够获得这些 用于实验的小鼠，但将在实验期间单独饲养动物 有时是漫长的领养迁移实验的过程。 这三个项目中的每一个都严重依赖于使用流式细胞术来 评估各种细胞表面标志物的表达。FACS的成本 分析包括在核心B中，因为：1)所有三个项目都将依赖于 大量使用共同的FACS设施；2)筛选 上述动物需要使用流式细胞术和3)许多 涉及流式细胞术的研究代表了协作交互作用 在私家侦探中。

The experiments proposed in this grant rely heavily on close collaborative interactions amongst the three PIs. The Core component will greatly facilitate these interactions. Core B is divided into 3 components: a) breeding of transgenic/knockout mice, b) maintenance of conventional transgenic and knockout mice, and c) flow cytometry. Core B is designed to provide aged T cell receptor transgenic, gene knockout and conventional mice that are required by all three projects of the program. Core B will breed and screen transgenic animals (using FACS or PCR analysis) and backcross them as necessary to render them suitable for the experiments detailed in the individual projects of the program. Because this process is very labor intensive, a centralized facility specialized to perform this task is essential. The core will obtain the various aged conventional, transgenic or knockout mice needed for the individual projects, quarantine them if necessary, develop standardized procedures for screening for the transgenic/knockout gene expression in each case, and will breed and backcross the mice to inbred strains. The goals are to have healthy, genetically homogeneous mice for the investigators in the project. Investigators will be able to obtain these mice for experiments but will maintain the animals separately during the course of the sometimes lengthy adoptive transfer experiments. Each of the three projects relies heavily on the use of flow cytometry to assess the expression of various cell surface markers. The costs for FACS analysis is included in Core B because: 1) all three projects will rely heavily on the use of the common FACS facility; 2) screening of the aforementioned animals require the use of flow cytometry and 3) many of the studies involving flow cytometry represent collaborative interactions among the PIs.