Onozuka Mari, Konno Hiroyuki, Akaji Kenichi, Nishino Hoyoku, Nosaka Kazuto
Department of Biochemistry and Molecular Biology, Kyoto Prefectural University of Medicine, Kamigyo-ku, Kyoto, Japan.
J Nutr Sci Vitaminol (Tokyo). 2005 Aug;51(4):274-7. doi: 10.3177/jnsv.51.274.
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 Sp1 site, specific DNA-protein complexes were identified. These findings indicate the importance of the Sp1 cis-element in regulating the hTPK1 gene expression.
我们克隆并分析了人硫胺素焦磷酸激酶基因（hTPK1）的5'-侧翼区域。使用瞬时转染的HepG2细胞进行的截短分析揭示了hTPK1启动子基础活性所需的最小区域，该区域编码在相对于转录起始位点的-105至+441之间的序列中。在使用来自HepG2细胞的核提取物和含有Sp1位点的合成寡核苷酸的电泳迁移率变动分析中，鉴定出了特异性的DNA-蛋白质复合物。这些发现表明Sp1顺式元件在调节hTPK1基因表达中的重要性。
