CORE--Gene Expression

主要研究者：Fisher Aron B。 a.核心单位名称 核心A：基因表达 名称eraCommons在项目中的作用 谢尔顿岛Feinstein博士范斯坦的核心主任宾夕法尼亚大学高级研究投资。 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物的研究。本申请中的所有拟议项目将研究表面活性剂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对这些项目至关重要。核心将为所有项目提供服务、工具、专门知识和援助，以实现这些目标。具体来说，核心将在E.大肠杆菌、昆虫细胞和 酵母巴斯德毕赤酵母进行定点诱变和缺失诱变， 构建敲入小鼠，对敲入、敲除或转基因小鼠进行基因型分析，提供实时PCR和Amaxa电穿孔系统用于转染培养物中的原代细胞的用途，以及制备和扩增腺病毒表达构建体。昆虫细胞和酵母表达系统、小鼠的PCR基因分型、敲入小鼠的构建体、实时PCR和Amaxa转染都是自2001年上一次计划更新以来核心增加的新服务。该计划中的所有项目都计划使用核心提供的服务。该中心还随时准备为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).