FRENCH Press

需要资金购买一台法国压榨机，以便高效、快速地从细菌、绿藻、酵母和植物细胞中提取蛋白质/酶。还要求提供两种型号的法国压力计，样品体积分别为3.7毫升和35毫升。这对细胞将满足我们相对多样化的蛋白质提取需求，范围从采集细胞的小样本(几克)到大样本(高达100克)。迫切需要这种设备来取代一台已有32年历史、大量使用的法国报刊，该报刊经常出现故障，目前无法运行。这让申请者和他们的HQP非常沮丧，他们的综合植物生物化学和分子生物学研究需要日常使用设备，有效和温和地提取在细菌细胞中异源表达的重组植物蛋白，或从绿藻、酵母或植物细胞培养物中提取天然蛋白。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，长时间的延误和随之而来的HQP生产率的降低正在经历之中。我们整合了一系列技术，并在很大程度上依赖于体外和体内的蛋白质生化分析，包括酶功能实验、蛋白质相互作用研究，以及关键蛋白质翻译后修饰的发生和作用的分析，如磷酸化、泛素化和糖基化。由一家法国出版社提供的分离具有生物活性的重组蛋白和天然蛋白的能力，对我们项目的成功绝对是必不可少的。我们的研究很好地促进了对鲜为人知的植物/藻类信号转导、胁迫反应和代谢途径的了解，并将在农业和生物燃料研究中具有深远的应用，这将有利于加拿大经济。这项建议的资助也将为我们的HQP提供一个重要的机会，使我们的HQP获得在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HQP培训和我们研究领域的合作，还将使申请者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取得具有更广泛影响和重要长期应用的关键发现。

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.