High capacity protein expression and production incubators

我的团队使用结构生物学，生物化学和生物信息学方法来研究莽草酸途径和密切相关的代谢途径如何驱动植物和微生物的功能多样性。莽草酸途径允许生物体适应其生长和发育的环境条件。该途径产生choris酸，这是芳香族化合物的前体，包括异黄酮、苯丙素、木质素、绿原酸、激素等。这些化合物在生物技术应用中具有重要作用。苯丙素、异黄酮类化合物和绿原酸具有强大的抗氧化特性，具有诱人的营养价值。莽草酸途径在动物中不存在，但在植物和微生物中是必不可少的，为抗菌剂和除草剂的开发提供了重要的酶靶点。因此，人们对研究这一途径中的蛋白质有着极大的兴趣。我们正在申请资金购买两台冷藏可堆叠式高容量培养摇床，型号为Innova S44i， ref 120V。这些可堆叠的孵化器摇床被要求取代我们的旧的和坏的孵化器摇床。我们的研究依靠冷藏摇床进行蛋白表达分析和重组蛋白的生产。具体来说，我们NSERC-DG资助的研究重点是蛋白质结构生物学和蛋白质生物化学，这使得纯化蛋白质成为我们研究中最有价值的货币。因此，推进我们的研究计划的能力直接取决于拥有可靠的冷藏孵化器振动器。蛋白质表达分析几乎完全在较低的温度下进行，在16 ？C和18 ？我们发现这个温度范围减缓了蛋白质翻译机制，从而减缓了蛋白质的生产，这使得蛋白质有足够的时间独立折叠而不形成包涵体。我们最初有两个冷藏培养摇床，一个用于蛋白质表达分析，另一个用于蛋白质制备，它们已经被我的研究小组和细胞与系统生物学系的其他植物和微生物研究实验室使用了20多年。然而，这两个孵化器在大流行期间停止了工作。非常幸运的是，我们能够从一个培养箱中抢救出零件来修复另一个。因此，我们的研究活动受到了影响，因为我们的冷藏培养箱摇床只有一个是正常工作的，它非常不可靠，急需更换。如果我们不能更换这些冷藏培养摇箱，我的NSERC-DG资助的研究项目和我的研究实验室的HQPs培训将进一步推迟，因为我的研究的所有方面都取决于我们成功获得重组蛋白的能力。

My group uses structural biology, biochemistry and bioinformatics approaches to investigate how the shikimate pathway and closely related metabolic pathways drive functional diversity in plants and microbes. The shikimate pathway allows organisms to adapt to environmental conditions for their growth and development. The pathway produces chorismate, a precursor for aromatic compounds including isoflavonoids, phenylpropanoids, lignin, chlorogenic acids, hormones etc. These compounds have important roles in biotechnological applications. Phenylpropanoids, isoflavonoids, and chlorogenic acids have powerful antioxidant properties with attractive nutritional benefits. The shikimate pathway is absent in animals but is essential in plants and microbes which provides important enzyme targets for the development of antimicrobials and herbicides. Therefore, there is tremendous interest in studying proteins from this pathway. We are requesting funding to purchase two refrigerated stackable high-capacity incubator shakers, model Innova S44i, ref 120V. These stackable incubators shakers are being requested to replace our old and broken incubator shakers. Our research depends on refrigerated shakers to conduct protein expression analysis and recombinant protein production. Specifically, our NSERC-DG funded research focuses on protein structural biology and protein biochemistry which makes purified proteins the most valuable currency in our studies. Therefore, the ability to advance our research program directly depends on having reliable refrigerated incubator shakers. Protein expression analysis are almost exclusively conducted at lower temperatures, between 16 ?C and 18 ?C. We found that this temperature range slows down the protein translation machinery and thus protein production, which allows sufficient time for proteins to independently fold without forming inclusion bodies. We initially had two refrigerated incubator shakers, one was designated for protein expression analysis and the other for protein preparation and they have been used for more than 20 years by my research group and other plant and microbial research labs in the Department of Cell and Systems Biology. However, both incubators stopped working during the pandemic. Quite fortunately we were able to salvage parts from one incubator to repair the other. As a result, our research activity has been impacted because only one of our refrigerated incubator shakers is functioning and it is highly unreliable and urgently needed to be replaced. If we are unable to replace these refrigerated incubator shakers my NSERC-DG funded research program and HQPs training in my research lab will be further delayed since all aspect of my research depends on our ability to successfully obtain recombinant proteins.