Microbial Production Core Facility

描述(由申请人提供)：我们建议购买New Brunswick Science BIOFLO5000 60升工作容量发酵罐和CEPA Z41大容量流动离心机，以建立主要用于表达重组蛋白的细菌和酵母菌株的微生物生产核心设施。拟议的核心设施将满足15名NIH支持的调查人员的需求，这些调查人员由34个不同的NIH奖项资助(22个R01、1个R21、1个R33、2个R37(功绩)、2个P01、1个P30、1个P41、1个S10、1个T32、1个U19、1个U54)。核心设施将为所有参与的实验室克服蛋白质纯化方面的重大瓶颈。参与这项应用的研究人员对130种不同的蛋白质进行了研究，这些蛋白质通常是野生型和突变型的，代表了总共200多种蛋白质制剂。计划中的核心设施在培养大型培养物方面比现有的可能性(10升发酵罐、摇瓶)更有效率，允许更大的蛋白质制剂，并将增强起始细胞材料的重复性。每个参与的研究项目都将大大节省时间。原料数量的飞跃将允许以足够的浓度提纯制备量的难溶蛋白质，最终允许对其进行生化和生物物理分析。我们选择New Brunswick Science BIOFLO5000和CEPA Z41是基于以前使用New Brunswick发酵罐和CEPA产品的经验、运行类似设施的个人的意见、定价和仪器的规格。相关性：本论文的研究人员从事基础和应用生物医学研究，使用纯化的蛋白质来确定与癌症和人类健康相关的机制。所要求的仪器不仅将使他们的工作显著提高效率，而且还将允许分析以前无法以足够数量或浓度纯化的蛋白质。

DESCRIPTION (provided by applicant): We propose to purchase a New Brunswick Scientific BIOFLO5000 60 liter working volume fermentor and a CEPA Z41 high capacity flow-through centrifuge to establish a Microbial Production Core Facility primarily for the growth of bacterial and yeast strains expressing recombinant proteins. The proposed core facility will serve the needs of 15 NIH-supported investigators, which are funded through 34 different NIH awards (22 R01, 1 R21, 1 R33, 2 R37 (MERIT), 2 P01, 1 P30, 1 P41, 1 S10, 1 T32, 1 U19, 1 U54). The core facility will overcome a significant bottleneck in protein purification for all laboratories involved. The investigators involved in this application work on 130 different proteins, often in their wild type and mutant forms, representing a total of over 200 protein preparations. The planned core facility is more efficient in growing large cultures than existing possibilities (10 liter fermentor, flasks), allows larger protein preparations, and will enhance the reproducibility of the starting cell material. Significant time savings for each participating research program will result. The leap in the amount of starting material will allow purifying preparative amounts of poorly soluble proteins at sufficient concentrations to finally allow their biochemical and biophysical analysis. Our choice of the New Brunswick Scientific BIOFLO5000 and the CEPA Z41 is based on previous experience with New Brunswick fermentors and CEPA products, the input from individuals running similar facilities, the pricing, and the specifications of the instruments. Relevance: The investigators of this submission engage in fundamental and applied biomedical research, using purified proteins to define mechanisms that are relevant to cancer and human health. The requested instrumentation will not only make their work significantly more efficient, but will also allow the analysis of proteins that previously could not be purified in sufficient quantities or concentrations.