Chemiluminescence detection system

本应用需要一种高分辨率的化学发光检测系统，用于使用化学发光和荧光对蛋白质(1D凝胶)、DNA和RNA的表达进行定量分析，并用于菌落计数。该设备应两名高级研究员(J.Lafond、D.Averill-Bates)和一名初级研究员(B.Barbeau)的要求，他们的NSERC研究项目都需要进行分子和代谢研究，需要对哺乳动物细胞和组织中蛋白质和核酸的表达进行定量分析。NSERC的研究计划旨在了解：(1)高胆固醇饮食对怀孕高胆固醇血症兔模型母体脂质代谢影响的信号机制；(2)来自人内源性逆转录病毒序列(HERV)的包膜蛋白在胎盘发育中的意义；以及(3)确定调控哺乳动物细胞中细胞应激反应(例如氧化应激)的细胞和分子机制。所要求的设备是最近的高分辨率(420万像素)技术，配备了高灵敏度的CCD相机，以及用于采集、分析和图像增强以及荧光和生物荧光成像的软件。这种设备在每个项目中都是必不可少的，用于通过Western blotting对细胞信号通路中涉及的多种蛋白质的表达、细胞内转位和磷酸化进行高灵敏度检测和定量分析。其他应用包括利用siRNA技术通过敲除后的Western blotts来定量蛋白质表达，或者通过再互补来过量表达。分子成像仪将是通过荧光标记的探针进行电泳法对细胞应激和氧化应激诱导的蛋白质损伤的mRNA和蛋白质表达变化进行量化的理想选择。购买高分辨率分子成像仪对于确保加拿大和国际层面的高质量和有竞争力的研究，以及NSERC资助的研究人员在生物化学、细胞和分子生物学方面培训至少100名研究生至关重要。

The present application requests a high-resolution Chemiluminescence detection system for the quantitative analysis of expression of proteins (1D gels), DNA and RNA using chemiluminescence and fluorescence, and for colony counting. The equipment is requested by 2 senior investigators (J. Lafond, D. Averill-Bates) and 1 junior investigator (B. Barbeau), whose NSERC research programs all entail molecular and metabolic investigations requiring quantitative analyses of expression of proteins and nucleic acids in mammalian cells and tissues. The NSERC research programs aim to understand: (1) the signalling mechanisms underlying the effects of a cholesterol-rich diet on metabolism of maternal lipids in a model of pregnant hypercholesterolemic rabbit; (2) the implication of envelope proteins derived from human endogenous retroviral sequence (HERV) in development of placenta; and (3) to identify the cellular and molecular mechanisms that regulate cellular stress responses (e.g. oxidative stress) in mammalian cells. The requested equipment is recent high-resolution (4.2 megapixels) technology that is equipped with a high sensitivity CCD camera, and software for acquisition, analysis and image enhancements, and for fluorescence and biofluorescence imaging. This equipment is essential in each of these projects for high-sensitivity detection and quantitative analysis of expression, intracellular translocation and phosphorylation of a multitude of proteins involved in cell signalling pathways, by Western blotting. Other applications include quantification of protein expression by Western blots following knock-down using siRNA technology or overexpression by recomplementation. The Molecular imager will be ideal for quantification of changes in mRNA and protein expression in response to cellular stress, and oxidative stress-induced protein damage by electrophoresis with fluorescently-labelled probes. The acquisition of a high-resolution Molecular imager is essential to ensure high quality and competitive research at the Canadian and international levels, and training of at least 100 graduate students by NSERC-funded investigators in Biochemistry, Cell and Molecular Biology.