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

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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.