Sensitive detection of protein-protein and nucleic acid-protein interactions using a digital imaging system

蛋白质-蛋白质相互作用的分子研究为许多细胞过程和途径提供了宝贵的启示，从细胞内信号的分子调节到对生物体发育和生命过程的大规模理解。同样，对蛋白质如何与核酸相互作用的研究重写了教科书，并彻底改变了我们对核酸的理解，无论是在核酸本身的水平上，还是在染色质结构和组织的更高水平上。从这些研究中获得的机械见解，照亮了我们对无数过程的理解，以及它们如何调节生命本身。重要的是，Pelka和Kumar小组的工作利用了这种分子生物学专业知识来探索调节宿主-病原体相互作用的途径和赋予抗生素耐药性的分子途径。我们nserc资助的研究重点是了解病毒蛋白如何在感染期间重新编程细胞网络以及细菌如何解除抗生素。这不仅使我们更清楚地了解了这些寄生虫超越宿主细胞的机制，而且还对宿主细胞水平和寄生虫本身的各种细胞调节网络和控制这些网络的机制提供了重要的见解。我们依靠老化的柯尼卡SRX 101-A显影剂和x射线胶片来满足我们的蛋白质、核酸和蛋白质核酸检测需求。这是我们nserc资助的许多研究成功的关键。如果没有检测蛋白质或核酸的能力，我们就无法完成NSERC发现基金的大部分目标。不幸的是，该系统太老了，柯尼卡不再支持它或制造备件，因为这样的维修和使用寿命是非常有限的，而更换零件是可用的。此外，经营胶片显影剂所需的耗材变得昂贵，而且更难获得；例如，显影剂和定影剂化学品在马尼托巴省已经没有了，我们从安大略省以更高的价格进口。同样，在过去的五年里，胶片的成本急剧上升，随着我们常规进行的western blots数量的增加，进行这些实验的费用也在飙升。再加上由于缺少化学品、薄膜或设备故障而导致的频繁停机，这极大地损害了我们执行某些类型的依赖于蛋白质或核酸敏感检测的检测的能力，同时损害了HQP的培训。最终，这个系统不可恢复的故障是迫在眉睫的。因此，我们迫切寻求用新的Azure 600数字成像系统取代我们过时的基于胶片的柯尼卡SRX 101-A，以继续我们的研究和HQP培训，而不必担心不可恢复的设备故障。至关重要的是，我们没有其他选择，我们的开发人员迫在眉睫的失败将剥夺我们对印迹进行成像的能力。

Molecular studies of protein-protein interactions have shed invaluable light on a multitude of cellular processes and pathways, ranging from the molecular regulation of intracellular signalling to large-scale understanding of organismal development and life processes. Likewise, studies of how proteins interact with nucleic acids have re-written textbooks and revolutionized our understanding of nucleic acids, both at the level of the nucleic acid itself and at the higher levels of chromatin structure and organization. Mechanistic insights gained from such studies have illuminated our understanding of countless processes and how they regulate life itself. Importantly, work carried out by the Pelka and Kumar groups have used such molecular biology expertise to probe pathways regulating host-pathogen interactions and molecular pathways conferring antibiotic resistance. Our NSERC-funded research focuses on the understanding of how viral proteins reprogram cellular networks during infection and how bacteria disarm antibiotics. This has led to a clearer understanding of not only the mechanisms by which these parasites overtake a host cell, but also have shed important insight into a variety of cellular regulatory networks and the mechanisms by which these networks are controlled, both at the level of the host cell and also within the parasite itself. We depend on an aging Konica SRX 101-A developer with X-ray film for our protein, nucleic acid, and protein-nucleic acid detection needs. It is the key to much of our NSERC-funded research success. Without the ability to detect proteins or nucleic acids we cannot complete the majority of our NSERC Discovery Grant goals. Unfortunately, the system is so old that Konica no longer supports it or manufactures spare parts, as such repairs and service lifetime is very limited while replacement parts are available. In addition, supplies necessary to operate the film developer have become expensive and more difficult to acquire; for example, the developer and fixer chemicals are no longer available in Manitoba and we import them from Ontario at greater cost. Likewise, the costs of film have risen dramatically over the last five years and with the number of western blots we routinely perform, the expense of carrying out these experiments have soared. Combined with frequent down-times due to lack of chemicals, film, or equipment breakdown, this has greatly compromised our ability to perform certain types of assays that rely on sensitive detection of proteins or nucleic acids while harming training of HQP. Ultimately, unrecoverable failure of this system is imminent. Thus, we urgently seek to replace our obsolete film-based Konica SRX 101-A with a new Azure 600 digital imaging system to continue our research and HQP training without fear of irrecoverable equipment failure. Crucially, there is no alternative for us to use and the looming failure of our developer will deprive us of the ability to image blots.