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.

该计划项目中涉及的三个实验室中的每个实验室中的每一个都将表达不同的蛋白质类别 从血清蛋白（APOAI）到这些子集的酶（LCAT和MPO）和突变形式，以便 如项目中所述进行随后的酶法测定，并利用多样化的分析和分析 在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 高体积蛋白表达的集中式平均值，从而促进了相对复杂的 以及关于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， 核心还将为所有三个项目提供分子克隆/诱变能力。核心 还将充当培训计划实验室成员的教育设施 使用各种策略以及在特定等位基因的突变体的产生中优化蛋白质表达 所需的蛋白质表达或转基因小鼠产生等位基因的靶向等位基因（CC/CWRU转基因 Mouse Core位于Case Western Reserve University）。培训蛋白质的纯化也将是 由核心提供。集中工作的这些方面和培训将大大提高 科学家参与每个项目的效率和生产力。