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、 和 3 个程序。该转基因动物核心的一个特殊优势是陈博士（核心领导者） 将为其他使用超氧化物歧化酶（SOD1、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 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）的原代细胞培养物中 啮齿类动物（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 用于对野生型原代细胞培养物进行设置、表征和质量控制的设施 动物，以及 SOD1 和 SOD2 转基因和敲除突变小鼠。此外，HSP70转基因 和 PARP-1 敲除小鼠将分别为项目 3 和项目 2 进行培育和基因分型 这个核心 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 氧化酶的酶活性和生化 在小鼠和原代细胞培养物中进行测定。最后，该核心将生成并表征双精度 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工具 使用各种转基因和基因敲除动物的目标。