| Research Abstract |
1. Cloning of promoter region of human mitochondrial phosphoenolpyruvate carboxykinase (PCK-2) gene
We screened 1.2x10^5 PAC clones by PCR, and obtained 2 positive clones, which contain the promoter region of PCK-2 gene. Analyzing these clones, we determined the nucleotide sequence up to about 1.9 kb upstream from start codon. The transient reporter assay using luciferase revealed that this region had significant promoter activity in human hepatoma cell line.
2. Analysis of hepatocyte nuclear factor-1α gene in Japanese diabetic patients.
We analyzed nucleotide sequence of hepatocyte nuclear factor-1α gene in Japanese diabetic patients, and determined two novel mutations of the gene.
3. Islet β-cells dysfunction by the over expression of gluconeogenic enzyme.
We established stable cell line expressing glucose-6-phosphatase from mouse β-cell clone MIN6. In this cell line, glucose responsive insulin release was decreased compared to the control cell line.
4. Analysis on the transcriptional regulations by insulin on rat PCK-1 gene.
We analyzed the transcriptional regulations by insulin on rat PCK-1 gene. The analysis revealed that the insulin signaling to the transcription factor FKHR might play some role in regulating PCK-1 gene by insulin, but that there should be another pathway.
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