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（由申请方提供）：需要一种发酵系统，包括用于细胞生长的中试规模发酵罐和用于细胞收获的连续流离心机。该系统将取代已有30年历史的现有设备，这些设备缺乏现代发酵罐的许多功能。斯坦福大学没有其他这样的发酵系统。它将用于生产蛋白质进行生物化学和结构研究，主要在四个研究领域，基因表达，细胞信号传导和免疫反应。基因表达的研究由Kornberg实验室进行转录，由Puglisi实验室进行蛋白质合成。Kornberg和他的同事们研究了来自酿酒酵母的RNA聚合酶II转录机制的结构和调节。几十年来，这项研究一直严重依赖于大规模发酵，以获得生物化学和晶体学所需的蛋白质数量。现在有人建议将整个努力付诸实践，为此目的需要一个现代发酵罐。Puglisi小组通过NMR和荧光方法研究原核生物中蛋白质合成的机制。需要大量的核糖体、辅助蛋白和tRNA。按要求的规模发酵将大大有助于这项工作。Weis及其同事研究细胞间相互作用、细胞信号传导和膜交易。他们目前正在研究通过表面受体的细胞粘附的分子基础，以及调节蛋白与负责细胞膜融合的SNARE的结合。所有这些工作都需要X射线晶体学分析，并依赖于细菌发酵。Wnt相互作用的研究特别需要低温细胞生长。Jardetzky小组研究病毒进入糖蛋白，IgE抗体和受体，TGF-B配体和受体。许多蛋白质在细菌中表达，并将受益于大规模细胞生长，最终通过X射线晶体学进行结构研究。 公共卫生相关性：拟议的发酵系统将使研究直接对基因表达，细胞信号传导和免疫反应的机制。在所提出的系统中大规模细胞培养物的生长将允许分离参与这些基本过程的蛋白质。将对分离的蛋白质进行生物化学和结构分析。