Fast Protein Liquid Chromatograhy System

这是一次重新申请资金，以购买GE Healthcare FPLC(快速蛋白质液体色谱)系统，该系统需要更换已有13年历史的FPLC，其中包含几个在过去3年中出现缺陷的关键部件。FPLC是我实验室中的关键设备，因为它几乎每天都被用于从植物组织和细胞培养中提纯和分析蛋白质催化剂(酶)，以及从转基因大肠杆菌中提取的蛋白质催化剂(酶)，在转基因大肠杆菌中，我们一直在研究的几种酶的野生型和突变版本已经过度表达。蛋白质纯化是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的关键酶的分子、调节和功能特性。我们的研究有助于推断植物如何改变和控制构成新陈代谢的化学反应，以响应不断变化的环境和/或发育参数。我们对通过磷酸化、单素化和糖基化进行的翻译后酶修饰特别感兴趣，因为这些修饰可以在控制酶活性、亚细胞定位、蛋白质：蛋白质相互作用和/或响应各种细胞外或细胞内信号的周转方面发挥关键作用。我们的研究对加拿大农业中的问题有着重要的长期应用，包括：(1)有针对性地修改油料种子(如油菜籽或大豆)中的储存油和蛋白质水平，(2)优化以植物为基础的大气二氧化碳到生物柴油和乙醇等可再生能源的转化，以及(3)开发磷高效作物，以减少人类对不可再生、不可持续和污染的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培训是不可能的。所要求的FPLC系统还将允许实验室人员利用过去十年开发的许多关于FPLC设计、计算机控制、色谱优化和数据采集的新技术。

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.