FRENCH Press

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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.