Transgenic Animal Core

该核心的目的是为项目1、2、 和3个节目。这个转基因动物核心的一个特别优势是陈博士(核心领导者) 将向其他使用超氧化物歧化酶(SO1、SOD2)的项目提供他的专业知识 转基因和基因敲除突变小鼠，以及NAD(P)H氧化酶小鼠(gp91Phox和p47Phox基因敲除) 如氧自由基相关的生物化学和分子生物学。此外，陈博士将提供他的专业知识 在原代培养神经元(方案2)、星形胶质细胞(方案3)和内皮细胞(方案1)中 啮齿动物(转基因、基因敲除和基因匹配的野生型)。该核心将作为一个集中的 野生型原代细胞培养的建立、鉴定和质量控制的设施 动物，以及SOD1和SOD2转基因和基因敲除突变小鼠。此外，HSP70转基因 将培育PARP-1基因敲除小鼠，并分别为项目3和项目2进行基因分型 还将提供基因表达的分子分析方面的专业知识(聚合酶链式反应、逆转录聚合酶链式反应、实时逆转录聚合酶链式反应)、 铜锌超氧化物歧化酶、锰超氧化物歧化酶、NAD(P)H氧化酶和生化的凝胶电泳法和酶活性 在小鼠和原代细胞培养中的检测。最后，这个核心将生成并描述替身 项目1的转基因/基因敲除小鼠(SOD1+/-/gp91Phox-/-，SOD2+/-/gp91Phox-/-，HSP70+/+/SOD2-/+) 3.因此，转基因动物核心为项目实现研究提供了独特的载体 目标是使用各种转基因和基因敲除动物。

The purpose of this Core is to make available various transgenic and knockout mutants for Projects 1, 2, and 3 of the program. One particular strength of this Transgenic Animal Core is that Dr. Chan (Core Leader) will provide his expertise to the other projects in the use of superoxide dismutases (SOD1, SOD2), in transgenic and knockout mutant mice, and NAD(P)H oxidase mice (gp91phox and p47phox knockout), as well as oxygen radical-related biochemistry and molecular biology. In addition, Dr. Chan will provide his expertise in primary cell culture of neurons (Project 2), astrocytes (Project 3), and endothelial cells (Project 1) from rodents (transgenic, knockout, and genetically-matched wild-types). This Core will serve as a centralized facility for set-up, for characterization, and for quality control of these primary cell cultures from wild-type animals, as well as SOD1 and SOD2 transgenic and knockout mutant mice. In addition, HSP70 transgenic and PARP-1 knockout mice will be bred and genotyped for Project 3 and Project 2, respectively This Core will also provide expertise in the molecular analysis of gene expression (PCR, RT-PCR, real-time RT-PCR), gel electrophoresis, and enzymatic activity of CuZnSOD, MnSOD, NAD(P)H oxidase, and biochemical assays in mice and in primary cell cultures. Finally, this Core will generate and characterize the double transgenic/knockout mice (SOD1+/- / gp91phox-/-, SOD2+/- / gp91phox-/-, HSP70+/+ / SOD2-/+) for Projects 1 and 3. Thus, the Transgenic Animal Core provides a unique vehicle for the projects to achieve the research goals employing various transgenic and knockout animals.