| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2011: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2010: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2009: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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| Research Abstract |
In order to clarify the transcriptional regulatory mechanisms of PIMT, isomerized protein repair enzyme, we cloned and functionally characterized the 5'-flanking(promoter) region of the gene. The putative promoter activity was confirmed using dual luciferase reporter gene assay system. The minimal region required for basal activity of the promoter was determined by generating a series of deletion and point mutation constructs. The binding protein(s) to the minimal region was briefly purified using biotin-labeled DNA probe, and subsequently LC-MS/MS analysis was performed and a transcriptional factor, NRF1, was identified. Binding activity of NRF1 to the minimal region was confirmed using electrophoretic mobility shift assay and chromatin immunoprecipitation assay.
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