FRENCH Press

需要资金购买一台法国压榨机，以便从细菌、绿色藻类、酵母和植物细胞中有效和迅速地提取蛋白质/酶。还要求提供两种型号的French Pressure Cell，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，从小（几克）到大（高达100克）的收获细胞样品提取。迫切需要这一设备来取代一台使用了32年、使用量很大的法国印刷机，该印刷机已变得容易频繁发生故障，目前无法操作。这对于申请人和他们的HQP来说是非常令人沮丧的，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来有效地和温和地提取在细菌细胞中异源表达的重组植物蛋白，或来自绿色藻类、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的细胞裂解，但由于持续的修复问题，HQP生产率正在经历长时间的延迟和随之而来的降低。我们整合了一系列的技术，并在很大程度上依赖于蛋白质生化分析，在体外和体内，包括酶功能实验，蛋白质-蛋白质相互作用的研究，并分析关键蛋白质翻译后修饰的发生和作用，如磷酸化，泛素化和糖基化。分离具有生物活性的重组蛋白和天然蛋白的能力，由法国出版社提供，对我们的计划取得成功至关重要。我们的研究是很好的定位，以推进知之甚少的植物/藻类信号转导，应激反应和代谢途径的知识，并将在农业和生物燃料的研究，将有利于加拿大经济具有深远的应用。这项建议的拨款亦会为我们的HQP提供一个重要的机会，让他们获得宝贵的培训，学习现代生化分析中广泛使用的重要技术。这不仅将扩大HQP培训和合作，在我们的研究领域，但也将定位申请人在关键发现，有更广泛的影响和重要的长期应用领域的植物驯化生物和非生物胁迫，信号转导和代谢途径的控制。

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.