Transgenic Animal Core

转基因动物核心

• 批准号: 7382861
• 负责人: PAK H CHAN
• 金额: $ 26.5万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7382861
• 关键词: Animals; Astrocytes; Biochemical; Biochemistry; Biological Assay; Breeding; Endothelial Cells; Gene Expression; Genotype; Goals; Heat-Shock Proteins 70; Knock-out; Knockout Mice; Molecular Analysis; Molecular Biology; Mus; Mutant Strains Mice; NAD(P)H oxidase; Neurons; Polymerase Chain Reaction; Primary Cell Cultures; Purpose; Quality Control; Reactive Oxygen Species; Research; Reverse Transcriptase Polymerase Chain Reaction; Rodent; SOD2 gene; Superoxide Dismutase; Time; Transgenic Animals; Transgenic Organisms; copper zinc superoxide dismutase; gel electrophoresis; knockout animal; mutant; programs

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.

该核心的目的是为项目1，2， 3、节目的。这个转基因动物核心的一个特别优势是，陈博士（核心领导人） 将提供他的专业知识，以其他项目在使用超氧化物歧化酶（SOD 1，SOD 2）， 转基因和敲除突变小鼠，以及NAD（P）H氧化酶小鼠（gp 91 phox和p47 phox敲除 与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， 在神经元（项目2）、星形胶质细胞（项目3）和内皮细胞（项目1）的原代细胞培养中， 啮齿类动物（转基因、敲除和遗传匹配的野生型）。这个核心将作为一个集中的 用于野生型原代细胞培养物的设置、表征和质量控制的设施 动物，以及SOD 1和SOD 2转基因和敲除突变小鼠。此外，HSP 70转基因 和PARP-1基因敲除小鼠将分别为项目3和项目2进行繁殖和基因分型。 还将提供基因表达分子分析（PCR、RT-PCR、实时RT-PCR）方面的专业知识， 凝胶电泳、CuZnSOD、MnSOD、NAD（P）H氧化酶和生化 在小鼠和原代细胞培养物中的测定。最后，这个核心将产生和表征双重 项目1的转基因/敲除小鼠（SOD 1 +/- /gp 91 phox-/-、SOD 2 +/- /gp 91 phox-/-、HSP 70 +/+ /SOD 2-/+） 和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 目的是利用各种转基因和基因敲除动物。