Role of cellular factors FUBP1 and DREF in regulation of viral and cellular gene expression via adenovirus E1A.

为了充分了解蛋白质的功能，特别是控制细胞内转录的蛋白质，需要进行一套专门的检测。这些检测包括染色质免疫沉淀检测蛋白质与细胞DNA的结合，荧光素酶检测检测启动子活性，实时定量PCR检测体内基因表达。这些试验是我提出的题为“细胞因子FUBP1和DREF在通过腺病毒E1A调节病毒和细胞基因表达中的作用”的发现基金的基石。该拨款中提出的实验需要专门的设备，我们正在向国家科学研究委员会申请资助。所要求的设备如下：用于染色质免疫沉淀测定的剪切染色质声纳- Covaris M220；能够处理384孔格式的实时PCR机- Bio-Rad CFX384，以及能够读取荧光素酶测定所需发光的多板读取器- Molecular Devices SpectraMax M3。该设备是根据几个标准选择的，包括功能集、易用性、耐用性、支持、保修和成本。这些设备对我们进行拟议研究的能力是必不可少的。如果没有所需的设备，我们的研究计划将受到严重影响，并将导致生产力和科学进步的降低。

In order to fully understand the function of proteins, especially proteins that control transcription within the cell, a specialized set of assays needs to be carried out. These assays include chromatin immunoprecipitation to look at protein binding to cellular DNA, luciferase assays to look at promoter activity and real-time quantitative PCR to look at gene expression in vivo. These assays are the corner stone of my proposed Discovery Grant titled "Role of cellular factors FUBP1 and DREF in regulation of viral and cellular gene expression via adenovirus E1A". The experiments proposed in that grant require specialized equipment, which we are requesting funding for from NSERC. The equipment requested is as follows: a sonicator for shearing chromatin for chromatin immuoprecipitation assays - Covaris M220; a real-time PCR machine capable of handling 384-well format - Bio-Rad CFX384, and a multi-plate reader capable of reading luminescence required for luciferase assays - Molecular Devices SpectraMax M3. This equipment was selected on the basis of several criteria, including feature-set, ease-of-use, durability, support, warranty and cost. The equipment is essential to our ability to carry out the proposed research. Without the requested equipment, our research programs will be drastically affected and will result in reduced productivity and scientific progress.