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

描述（由申请人提供）：要求一种发酵系统，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动离心机。该系统将取代已有30年历史的现有设备，并且缺乏现代发酵罐的许多功能。斯坦福大学没有其他类似的发酵系统。它将用于生产用于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和他的同事研究了酿酒酵母RNA聚合酶II转录机制的结构和调控。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。现在有人建议把所有的努力付诸实践，为此需要一台现代化的发酵罐。Puglisi小组利用核磁共振和荧光方法研究了原核生物蛋白质合成的机制。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方便工作。Weis和他的同事研究细胞-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研究细胞粘附的分子基础，以及调节蛋白与负责细胞膜融合的SNAREs的结合。所有这些工作都需要x射线晶体分析，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，以及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。

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.