|Budget Amount *help
¥107,640,000 (Direct Cost: ¥82,800,000、Indirect Cost: ¥24,840,000)
Fiscal Year 2011: ¥18,330,000 (Direct Cost: ¥14,100,000、Indirect Cost: ¥4,230,000)
Fiscal Year 2010: ¥18,330,000 (Direct Cost: ¥14,100,000、Indirect Cost: ¥4,230,000)
Fiscal Year 2009: ¥21,060,000 (Direct Cost: ¥16,200,000、Indirect Cost: ¥4,860,000)
Fiscal Year 2008: ¥22,620,000 (Direct Cost: ¥17,400,000、Indirect Cost: ¥5,220,000)
Fiscal Year 2007: ¥27,300,000 (Direct Cost: ¥21,000,000、Indirect Cost: ¥6,300,000)
To develop the system to study gene function at tissue level in medaka, we have established two methods in the present study. 1) Reverse genetics : TILLING(Targeting Induced Local Lesions IN Genome) is a reverse genetic strategy that combines random chemical mutagenesis with high-throughput discovery of induced mutations in the target genes. The method has been applied to a variety of plant and animal species. Screening of induced mutation is the most important step in TILLING. We developed the High Resolution Melting(HRM) assay, evaluated its efficacy for screening ENU-induced mutations in the medaka TILLING library. The HRM assay is fast, cost-effective, and robust in detecting variations in DNA. By HRM assay many nonsense mutations were identified and the phenotypes of these nonsense mutants confirmed their loss-of-function nature. 2) Regulation of gene expression in tissue : Infrared laser. evoked gene operator(IR-LEGO) is a microscope system optimized for heating cells at single cell level. Heat stress induces transcription of genes that are under the control of a heat shock promoter in individual targeted cells. We applied this technique to medaka fish carrying a heat shock promoter driven transgene and have succeeded the control of ectopic gene expression.