Molecular Cloning and Analysis of the 5'-Flanking Region of the Human Thiamin Pyrophosphokinase Gene

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  • ONOZUKA Mari
    Department of Biochemistry and Molecular Biology, Kyoto Prefectural University of Medicine
  • KONNO Hiroyuki
    Department of Chemistry, Kyoto Prefectural University of Medicine
  • AKAJI Kenichi
    Department of Chemistry, Kyoto Prefectural University of Medicine
  • NISHINO Hoyoku
    Department of Biochemistry and Molecular Biology, Kyoto Prefectural University of Medicine
  • NOSAKA Kazuto
    Department of Chemistry, Kyoto Prefectural University of Medicine

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We cloned and analyzed the 5'-flanking region of the human thiamin pyrophosphokinase gene (hTPK1). Truncation analysis using transiently transfected HepG2 cells revealed the minimal region required for basal activity of the hTPK1 promoter, which was encoded in a sequence between -105 and +441 relative to the transcription start site. In an electrophoretic mobility shift assay using the nuclear extracts from HepG2 cells and the synthetic oligonucleotide containing the Sp 1 site, specific DNA-protein complexes were identified. These findings indicate the importance of the Spl cis-element in regulating the hTPK1 gene expression.

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