CORE--Gene Expression

主要调查者：费舍尔·阿伦·B。 A.核心单位职称 核心A：基因表达 名称eraCommons在项目从属关系职位上的角色 谢尔顿·I·范斯坦，费因斯坦大学核心主任博士。老宾夕法尼亚研究投资公司。 摘要： 基因表达核心，核心A的目的是促进对项目感兴趣的基因和基因产品的研究。所有在这项申请中提出的项目都将研究表面活性物质代谢中的关键蛋白质。在体外、细胞和动物中研究这些蛋白质的能力将是这些项目的关键。核心将向所有项目提供服务、工具、专门知识和援助，以实现这些目标。具体地说，Core将在大肠杆菌、昆虫细胞和 酵母，毕赤氏酵母，进行定点突变和缺失突变，协助 构建敲入小鼠，对敲入、敲除或转基因小鼠进行基因分析，利用实时聚合酶链式反应和Amaxa电穿孔系统将原代细胞转入培养中，并制备和扩增腺病毒表达载体。昆虫细胞和酵母表达系统、小鼠的聚合酶链式反应基因分型、转基因小鼠的构建、实时聚合酶链式反应和Amaxa的转基因都是Core自2001年上一次计划更新以来增加的新服务。该计划中的所有项目都计划使用Core提供的服务。核心还随时准备在分子生物学和基因表达的其他领域提供援助，例如使用小干扰RNA(SiRNA)。

Principal Investigator: Fisher Aron B. a. TITLE OF CORE UNIT CORE A: GENE EXPRESSION Name eraCommons Role on Project Affiliation Position Sheldon I. Feinstein, Ph.D. feinsteins Core Director Univ. Penn Sr. Research Invest. ABSTRACT: The purpose of the Gene Expression Core, Core A is to facilitate the study of the genes and gene products that are of interest to the projects. All of the proposed projects in this application will examine key proteins in surfactant metabolism. The ability to study these proteins in vitro, in cells and in animals will be critical to the projects. The Core will provide services, instruments, expertise and assistance to all the projects in accomplishing these goals. Specifically, the Core will generate recombinant proteins in E. coli, insect cells and the yeast, Pichia pastoris, perform site-directed mutagenesis and deletion mutagenesis, assist in the construction of knock-in mice, perform genotype analysis for knock-in, knockout or transgenic mice, provide use of a real-time PCR and the Amaxa electroporation system for transfections of primary cells in culture and prepare and amplify adenoviral expression constructs. The insect cell and yeast expression systems, the PCR genotyping of mice, the constructs for knock-in mice, the real-time PCR and the transfection with Amaxa are all new services added by the Core since the last program renewal in 2001. All of the projects in the Program plan to use services offered by the Core. The Core also stands ready to provide assistance with other areas of molecular biology and gene expression such as use of small interfering RNA (siRNA).