Stress tolerance : gel documentation for analysis of gene/protein/enzyme expression and adaptation

压力耐受性：用于分析基因/蛋白质/酶表达和适应的凝胶文档

• 批准号: 375674-2009
• 负责人: Storey, Kenneth
• 金额: $ 3.05万
• 依托单位: Carleton University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments - Category 1 (<$150,000)
• 财政年份: 2008
• 资助国家: 加拿大
• 起止时间: 2008-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=412200
• 关键词: Stress; tolerance; gel; documentation; analysis

Research in the Storey lab at Carleton University explores the biochemical adaptations that allow animals to endure environmental extremes. We explore amazing natural phenomena including hibernation, freeze tolerance, cold hardiness, estivation and anoxia tolerance; in particular, our research on the biochemistry of winter survival by Canadian animals is making unique discoveries that are world-leading. All of these phenomena are linked by their use of torpor or dormancy to provide life extension under harsh conditions and we have identified multiple mechanisms that coordinate entry into hypometabolic states, reprioritize metabolic energy use, and preserve/stabilize cells and macromolecules to provide long term viability during dormancy. This research not only identifies key principles and methods of metabolic regulation that underlie environmental stress adaptation across the animal kingdom but it has important applications in biomedicine because methods for inducible hypometabolism could greatly improve procedures for storage of human tissues and organs removed for transplantation. The work of all researchers in my lab relies heavily on methods of gel electrophoresis for separation of proteins and DNA followed by the use of gel documentation instrumentation for image acquisition and data quantification. All lab members (typically 15-20 researchers) use gel documentation to analyze gene up-regulation, transcription factor activation, protein expression, enzyme purity, or protein/enzyme phosphorylation states. The entire lab relied on one gel documentation system and this aging machine was not only experiencing recurring expensive breakdowns but the technology had become a rate-limiting bottleneck in the lab because it was needed on a daily basis by most researchers. Then disaster struck in July 2008 with the catastrophic failure of the camera unit which halted all research and is still not repaired as of October 2008. Our lab presently relies on a "loaner" piece of equipment. The current application seeks to purchase the G:Box Chemi XT16 gel imaging system (Syngene Corp) to replace our current machine and provide state-of-the-art, high resolution gel imaging and quantification to support all of our research goals.

卡尔顿大学Storey实验室的研究探索了使动物能够忍受极端环境的生物化学适应性。我们探索令人惊叹的自然现象，包括冬眠，抗冻性，耐寒性，夏眠和耐缺氧性;特别是，我们对加拿大动物冬季生存的生物化学研究正在取得世界领先的独特发现。所有这些现象都通过它们在恶劣条件下使用休眠或休眠来提供寿命延长而联系在一起，并且我们已经确定了多种机制，这些机制协调进入低代谢状态，重新优先考虑代谢能量使用，并保存/稳定细胞和大分子以在休眠期间提供长期活力。这项研究不仅确定了动物界环境应激适应的代谢调节的关键原则和方法，而且在生物医学中具有重要的应用，因为诱导性低代谢的方法可以大大改善用于移植的人体组织和器官的储存程序。我实验室的所有研究人员的工作在很大程度上依赖于凝胶电泳分离蛋白质和DNA的方法，然后使用凝胶记录仪器进行图像采集和数据定量。所有实验室成员（通常15-20名研究人员）使用凝胶文件来分析基因上调，转录因子激活，蛋白质表达，酶纯度或蛋白质/酶磷酸化状态。整个实验室依赖于一个凝胶记录系统，这个老化的机器不仅经常出现昂贵的故障，而且该技术已经成为实验室的限速瓶颈，因为大多数研究人员每天都需要它。2008年7月，相机单元发生灾难性故障，停止了所有研究，截至2008年10月仍未修复。我们的实验室目前依靠一台“借来的”设备。目前的申请旨在购买G：Box Chemi XT 16凝胶成像系统（Syngene Corp），以取代我们目前的机器，并提供最先进的高分辨率凝胶成像和定量，以支持我们所有的研究目标。