FRENCH Press

法国媒体

• 批准号: RTI-2017-00060
• 负责人: Plaxton, William
• 金额: $ 2.9万
• 依托单位: Queen's University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2016
• 资助国家: 加拿大
• 起止时间: 2016-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=616536
• 关键词: FRENCH; Press

Funds are requested to purchase a French Press for efficient and rapid protein/enzyme extraction from bacterial, green algal, yeast, and plant cells. Also requested are two models of French Pressure Cells with respective sample volumes of 3.7 and 35 mL. This pair of Cells will satisfy our relatively diverse protein extraction needs which range from extractions from small (several grams) to large (up to 100 gram) samples of harvested cells. This equipment is urgently needed to replace a 32 year-old and heavily-used French Press that has become prone to frequent breakdowns and is currently inoperable. This has been very frustrating for the applicants and their HQP whose integrated plant biochemistry and molecular biology research requires daily access to equipment that effectively and gently extracts recombinant plant proteins that have been heterologously expressed in bacterial cells, or native proteins from green algal, yeast, or plant cell cultures. Virtually all of our collective research projects rely on large-scale cell lysis, but long delays and the consequent reduction in HQP productivity are being experienced due to on-going repair issues. We integrate a range of techniques and rely heavily on protein biochemical assays, both in vitro and in vivo, including enzyme function experiments, protein-protein interaction studies, and analysis of the occurrence and roles of crucial protein post-translational modifications such as phosphorylation, ubiquitination, and glycosylation. The ability to isolate biologically active recombinant and native proteins, enabled by a French Press, is absolutely essential to the success of our programs. Our research is well positioned to advance knowledge of poorly understood plant/algal signal transduction, stress responses, and metabolic pathways and will have far-reaching applications in agricultural and biofuels research that will benefit the Canadian economy. Funding of this proposal will also provide an important opportunity for our HQP to gain valuable training with a widely used and important technique in modern biochemical analyses. This will not only expand HQP training and collaborations in our areas of research, but will also position the applicants in making key discoveries that have broader implications and important long-term applications in the fields of plant acclimation to biotic and abiotic stress, signal transduction, and the control of metabolic pathways.

需要资金购买法国压榨机，以便从细菌、绿藻、酵母和植物细胞中高效快速地提取蛋白质/酶。还要求提供两种型号的法国压力传感器，样品体积分别为 3.7 和 35 mL。这对细胞将满足我们相对多样化的蛋白质提取需求，范围从小量（几克）到大量（最多 100 克）收获细胞样品的提取。急需该设备来替换一台已有 32 年历史且使用频繁的法式压滤机，该压滤机经常出现故障，目前无法运行。这对于申请人及其总部来说非常令人沮丧，他们的综合植物生物化学和分子生物学研究需要每天使用设备，有效、温和地提取在细菌细胞中异源表达的重组植物蛋白，或来自绿藻、酵母或植物细胞培养物的天然蛋白。事实上，我们所有的集体研究项目都依赖于大规模细胞裂解，但由于持续的修复问题，导致了长时间的延误以及随之而来的 HQP 生产力的下降。我们整合了一系列技术，并严重依赖体外和体内的蛋白质生化检测，包括酶功能实验、蛋白质-蛋白质相互作用研究，以及关键蛋白质翻译后修饰（如磷酸化、泛素化和糖基化）的发生和作用分析。通过法式压滤机分离具有生物活性的重组蛋白和天然蛋白的能力对于我们项目的成功绝对至关重要。我们的研究有利于增进人们对植物/藻类信号转导、应激反应和代谢途径知之甚少的了解，并将在农业和生物燃料研究中产生深远的应用，从而有利于加拿大经济。该提案的资助还将为我们的总部提供一个重要的机会，让他们获得现代生化分析中广泛使用的重要技术的宝贵培训。这不仅将扩大我们研究领域的 HQP 培训和合作，而且还将使申请人能够在植物适应生物和非生物胁迫、信号转导和代谢途径控制等领域做出具有更广泛影响和重要长期应用的关键发现。