Core B- Expression

这一核心的目的是为该方案的四个项目提供所需的细胞和蛋白质。这个 核心B人员要分离的特定细胞是：1)血细胞，主要是人和小鼠的血小板 来源；2)人和小鼠来源的内皮细胞；以及3)真核或原核细胞表达某些特定的 整合素、细胞骨架蛋白及其突变体。原核细胞被用作 纯化的蛋白质；talin，kindlins和scr家族的激酶衍生物，在核心中，并提供给个体 根据需要进行项目。由经验丰富的人员制备细胞将有助于确保纯度和 功能。特别是要有项目所需的可靠的功能性血小板制剂来源 有价值的，因为血小板的功能可能是高度可变的。小鼠血管内皮细胞也是如此 实验进展通常依赖于对来自于 少量有价值的小鼠品系。纯化的蛋白质用于结合研究和结构分析。 分析，需要相对大量的高质量制剂。需要准备一份 蛋白质数量有限，分离程序可以标准化，提供高质量的产品。至 集中执行这些活动将节省资源，有助于确保质量，并允许项目间 结果比较。此外，由于个别项目仍将需要相当大的独立性 细胞培养和分离细胞无疑将需要进行特定的实验，核心将在训练中发挥作用 该计划项目的成员在适当的细胞培养和分离方法以及 适当的方法来评估细胞的质量。因此，核心也起到了教育作用。

The purpose of this core is to provide cells and proteins as needed to the four Projects of the Program. The specific cells to be isolated by Core B personnel are: 1) blood cells, primarily platelets of human and murine origin; 2) endothelial cells of human and murine origin; and 3) eukaryotic or prokaryotic cells expressing certain integrins, cytoskeletal proteins and mutants of these proteins. The prokaryocytic cells are used as a source for purified proteins; talin, kindlins and scr-family kinase derivatives, in the core and provide to the individual projects as needed. The preparation of cells by experience personnel will help to insure uniformity of purity and function. To have a reliable source of functional platelet preparations as needed by the projects is particularly valuable since the function of platelets can be highly variable. This is also true of endothelial cells of murine origin where experimental progress often depends on performing experiments with the cells derived from a small number of valuable mouse strains. The purified proteins are used in binding studies and for structural analyses, where relatively large quantities of high quality preparations are needed. With the need to prepare a limited number of proteins, isolation procedures can be standardized to provide high quality products. To perform these activities centrally will conserve resources, will help to insure quality and will allow for interproject comparison of results. In addition, since the individual Projects will still require substantial independent cell culture and will undoubtedly need to isolate cells for specific experiments, the Core will function in training members of the Program Project laboratories in proper cell culture and isolation methodologies and in appropriate ways to assess the quality of the cells. Thus, the Core serves an educational function as well.