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种类的蛋白质 从血清蛋白（ApoAI）到酶（LCAT和MPO）以及这些酶的子集的突变形式， 按照项目中所述进行后续酶测定，并利用各种 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的是提供一个 用于高容量蛋白质表达的集中式装置， 以及对每个项目感兴趣的蛋白质进行的各种生物物理和生物学研究，此外， 核心还将为所有三个项目提供分子克隆/诱变能力。核心 还将作为一个教育设施，培训计划项目实验室的成员， 使用各种策略优化蛋白质表达，以及产生特定等位基因的突变体 对于蛋白质表达或用于靶向等位基因以产生转基因小鼠（CC/CWRU转基因小鼠）是期望的 小鼠核心位于凯斯西储大学）。蛋白质纯化方面的培训也将在 由核心提供。将这些方面的工作和培训集中起来，预计将大大加强 参与每个项目的科学家的效率和生产力。