Recombinant Protein Expression and Molecular Cloning

重组蛋白表达和分子克隆

• 批准号: 8015699
• 负责人: Joseph DiDonato
• 金额: $ 15.8万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-08 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8015699
• 关键词: Alleles; Biological; Biological Assay; Biophysics; Cloning; Educational process of instructing; Enzymes; Generations; Human Resources; Knock-in Mouse; Laboratories; Mass Spectrum Analysis; Molecular Cloning; Mutagenesis; Phosphatidylcholine-Sterol O-Acyltransferase; Production; Productivity; Proteins; Recombinant Proteins; Research Personnel; Scientist; Serum Proteins; Techniques; Training; Transgenic Mice; Transgenic Organisms; Universities; Work; experience; expression cloning; instrumentation; interest; meetings; member; mutant; programs; protein expression; protein purification

Each of the three laboratories involved in the Program Project will express distinct classes of proteins from serum proteins (ApoAI) to enzymes (LCAT, and MPO) and mutant forms of a subset of these in order to perform subsequent enzymatic assays as described in the Projects and analysis utilizing diverse and sophisticated instrumentation detailed in the Mass Spectrometry and Biophysics Core (Core B). The principal objective ofthe Recombinant Protein Expression and Molecular Cloning Core is to provide a centralized means for high volume protein expression thereby facilitating the relatively sophisticated and diverse biophysical and biological studies on the proteins of interest for each Project, in addition, the Core will also provide molecular cloning/ mutagenesis capabilities to all three Projects. The Core will also function as an educational facility for training members of the Program Project laboratories on ways to optimize protein expression using various strategies and also in the generation of mutants of specific alleles desired for protein expression or for targeting alleles for transgenic mouse production ( CC/CWRU Transgenic mouse Core is located at Case Western Reserve University). Training in the purification of proteins will also be provided by the Core. Centralizing these aspects of the work and training is expected to greatly enhance efficiency and productivity of the scientists participating in each project.

参与该计划项目的三个实验室中的每一个都将表达不同类别的蛋白质 从血清蛋白（ApoAI）到酶（LCAT 和 MPO）以及其中一部分的突变形式，以便 使用各种和分析进行项目和分析中描述的后续酶测定 质谱和生物物理学核心（核心 B）中详细介绍了复杂的仪器。校长 重组蛋白表达和分子克隆核心的目标是提供 用于大量蛋白质表达的集中手段，从而促进相对复杂的 以及针对每个项目感兴趣的蛋白质的各种生物物理和生物学研究，此外， 该核心还将为所有三个项目提供分子克隆/诱变能力。核心 还将作为教育设施，对计划项目实验室的成员进行培训 使用各种策略以及特定等位基因突变体的生成来优化蛋白质表达 蛋白质表达或转基因小鼠生产的靶向等位基因所需的（CC/CWRU Transgenic mouse Core 位于凯斯西储大学）。蛋白质纯化方面的培训也将进行 由核心提供。集中这些方面的工作和培训预计将大大提高 参与每个项目的科学家的效率和生产力。