Facilitating large scale biomass generation: acquisition of an automated 100 L fe

促进大规模生物质发电：购买自动化 100 L fe

• 批准号: 7795468
• 负责人: ROGER D KORNBERG
• 金额: $ 32.38万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-12-01 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7795468
• 关键词: Area; Bacteria; Biochemical; Biochemistry; Biomass; Cell Adhesion; Cell Communication; Cell Culture Techniques; Cell membrane; Cells; Crystallography; Equipment; Fc Receptor; Fermentation; Fluorescence; Gene Expression; Generations; Genetic Transcription; Glycoproteins; Growth; Harvest; IgE; Immune response; Laboratories; Ligands; Membrane; Membrane Fusion; Methods; Molecular; Process; Production; Prokaryotic Cells; Protein Biosynthesis; Proteins; RNA Polymerase II; Regulation; Research; Ribosomes; Roentgen Rays; Saccharomyces cerevisiae; Signal Transduction; Structure; Surface; System; Universities; Viral; Work; X-Ray Crystallography; Yeasts; base; cell growth; cold temperature; genetic regulatory protein; public health relevance; receptor; receptor binding

DESCRIPTION (provided by applicant): A fermentation system is requested, comprising a pilot scale fermentor for cell growth and a continuous flow centrifuge for cell harvest. This system will replace existing equipment that is 30 years old and lacks many capabilities of modern fermentors. There is no other such fermentation system available at Stanford University. It will be used for the production of proteins for biochemical and structural studies, primarily in four research areas, gene expression, cell signaling, and the immune response. Studies of gene expression are performed by the Kornberg laboratory on transcription and by the Puglisi laboratory on protein synthesis. Kornberg and colleagues investigate the structure and regulation of the RNA polymerase II transcription machinery from the yeast Saccharomyces cerevisiae. This research has been critically dependent for several decades on large scale fermentation, to obtain the quantities of proteins needed for biochemistry and crystallography. It is now proposed to bring the entire effort to fruition, and a modern fermentor is needed for the purpose. The Puglisi group study mechanisms of protein synthesis in prokaryotes by NMR and fluorescence methods. Large amounts of ribosomes, accessory proteins, and tRNAs are required. Fermentation on the scale requested will greatly facilitate the work. Weis and colleagues study cell-cell interaction, cell signaling, and membrane transactions. They are currently pursuing the molecular basis of cell adhesion through the surface receptor, and binding of regulatory proteins to the SNAREs responsible for cell membrane fusion. All of this work entails X-rays crystallographic analysis, and depends on bacterial fermentation. The studies of Wnt interaction particularly require low temperature cell growth. The Jardetzky group study viral entry glycoproteins, IgE antibodies and receptors, and TGF-B ligands and receptors. Many of the proteins are expressed in bacteria, and will benefit from large scale cell growth for eventual structural studies by X-ray crystallography. PUBLIC HEALTH RELEVANCE: The proposed fermentation system will enable research directed towards the mechanisms of gene expression, cell signaling, and the immune response. Growth of cell cultures on a large scale in the proposed system will permit the isolation of proteins involved in these fundamental processes. The isolated proteins will be subjected to biochemical and structural analyses.

描述（由申请人提供）：请求发酵系统，包括用于细胞生长的试验量表发酵罐和用于细胞收获的连续流动离心机。该系统将取代30岁的现有设备，并且缺乏现代发酵罐的许多功能。斯坦福大学没有其他此类发酵系统可用。它将用于生产生化和结构研究的蛋白质，主要用于四个研究领域，基因表达，细胞信号传导和免疫反应。基因表达的研究是由Kornberg实验室在转录和Puglisi实验室的蛋白质合成方面进行的。 Kornberg及其同事研究了酿酒酵母的RNA聚合酶II转录机制的结构和调节。几十年来，这项研究一直依赖于大规模发酵，以获取生物化学和晶体学所需的蛋白质数量。现在建议将全部努力带来实现，为此目的需要现代发酵罐。 NMR和荧光方法中蛋白质合成的Puglisi组研究机制。需要大量核糖体，辅助蛋白和TRNA。所要求的规模的发酵将极大地促进工作。 Weis及其同事研究细胞 - 细胞相互作用，细胞信号传导和膜交易。他们目前正在通过表面受体追求细胞粘附的分子基础，并结合了调节蛋白与负责细胞膜融合的鼻涕的结合。所有这些工作都需要X射线晶体学分析，并取决于细菌发酵。 Wnt相互作用的研究特别需要低温细胞生长。 Jardetzky组研究病毒进入糖蛋白，IgE抗体和受体以及TGF-B配体和受体。许多蛋白质在细菌中表达，将受益于大规模细胞的生长，用于通过X射线晶体学最终的结构研究。 公共卫生相关性：拟议的发酵系统将使研究针对基因表达，细胞信号传导和免疫反应的机制。拟议系统中大规模的细胞培养物的生长将允许隔离这些基本过程中涉及的蛋白质。分离的蛋白质将进行生化和结构分析。