Fast Protein Liquid Chromatograhy System

这是购买GE Healthcare FPLC（快速蛋白质液相色谱）系统的资金重新申请，需要更换13年的FPLC，其中包含过去3年中出现缺陷的几个重要组件。FPLC是我实验室的关键设备，因为它几乎每天都用于从植物组织和细胞培养物以及转基因大肠杆菌提取物中纯化和分析蛋白催化剂（酶）。我们研究的几种酶的野生型和突变体在大肠杆菌中过表达。蛋白质纯化形成了我实验室大部分研究的基础，因为我们的长期目标是阐明植物碳水化合物和磷酸盐代谢关键酶的分子，调节和功能特性。我们的研究有助于推断植物如何修改和控制构成代谢的化学反应，以应对不断变化的环境和/或发育参数。我们特别感兴趣的是通过磷酸化，monoubiquitination和糖基化的翻译后酶修饰，因为这些可以在控制酶活性，亚细胞位置，蛋白质：蛋白质相互作用和/或营业额响应于各种细胞外或细胞内信号中发挥关键作用。我们的研究对加拿大农业问题有重要的长期应用，包括：（1）有针对性地改变含油种子（如油菜或大豆）中的储存油与蛋白质水平，（2）优化基于植物的大气CO2向可再生能源（如生物柴油和乙醇）的转化，以及（3）开发磷酸盐高效作物，迫切需要减少人类对不可再生、不可持续和污染性磷肥的猖獗但低效的使用。如果没有一流的FPLC系统，这项研究和相关的HQP培训将是不可能的。所要求的FPLC系统还将允许实验室人员利用在过去十年中开发的关于FPLC设计、计算机控制、色谱优化和数据采集的许多新技术。

This is a reapplication for funds to purchase a GE Healthcare FPLC (Fast Protein Liquid Chromatography) system, needed to replace a 13-year-old FPLC containing several vital components that have become defective over the past 3 years. The FPLC is a pivotal piece of equipment in my lab since it is used virtually on a daily basis for the purification and analysis of protein catalysts (enzymes) from plant tissues and cell cultures, as well as from extracts of transgenic E. coli in which wild-type and mutant versions of several enzymes that we have been studying have been overexpressed. Protein purification forms the foundation upon which the majority of my lab's research is built since our long-term goal is to elucidate the molecular, regulatory, and functional properties of key enzymes of plant carbohydrate and phosphate metabolism. Our research helps to deduce how plants modify and control the chemical reactions that constitute metabolism in response to changing environmental and/or developmental parameters. We are particularly interested in post-translational enzyme modifications by phosphorylation, monoubiquitination, and glycosylation since these can play pivotal roles in controlling enzyme activity, subcellular location, protein:protein interactions and/or turnover in response to various extra- or intracellular signals. Our research has significant long-term applications to problems in Canadian agriculture including the: (1) targeted modification of storage oil versus protein levels in oilseeds such as canola or soybean, (2) optimizing plant-based conversion of atmospheric CO2 into renewable energy sources such as biodiesel and ethanol, and (3) development of phosphate-efficient crops, urgently needed to reduce mankind's rampant but inefficient use of non-renewable, unsustainable, and polluting phosphate fertilizers. This research and associated HQP training would be impossible without ready access to a top-notch FPLC system. The requested FPLC system would also allow lab personnel to exploit many new technologies regarding FPLC design, computer control, chromatographic optimization, and data acquisition that have been developed over the past decade.