Deletion analysis of the promoter of Aspergillus oryzae gene encoding heat shock protein 30(GENETICS, MOLECULAR BIOLOGY, AND GENE ENGINEERING)
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概要
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In order to find a promoter that could be influenced by temperature shift, we explored and isolated an Aspergillus oryzae gene expressed at high temperatures (37-42℃) by the cDNA subtraction method. Of the 96 cDNA clones isolated from the subtraction library, one cDNA clone showed 73% identity with Aspergillus nidulans heat shock protein 30 (hsp30). Based on this, we designated the isolated gene hsp30 of A. oryzae. A. oryzae hsp30 was weakly expressed at 30℃, but strongly at 40℃. We showed that the promoter of this hsp30 induced heterologous gene expression at high temperatures using β-glucuronidase (GUS) gene as a reporter. Regarding elucidation of the region essential for heat shock response, we showed that the minimum length of the promoter region that was essential for heat shock response was located between -388 and -272 (+1 indicated the first position of the translation initiation codon) of the hsp30 promoter. This promoter region harbors several putative transcription factor binding sites, including heat shock elements (HSEs), a CCAAT box, and a TATA box. Furthermore, site-directed mutagenesis of this promoter revealed that HSE1 (aTTCgtcGAAacgcccaGAAa) and HSE2 (cGAAagTTCtcGACg), located between -342 and -272 of the hsp30 promoter, were its cis-acting elements for heat shock response.
著者
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Kashiwagi Yutaka
National Food Research Institute
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TADA Sawaki
National Food Research Institute, National Agriculture and Food Research Organization (NARO)
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SUZUKI Satoshi
National Food Research Institute, National Agriculture and Food Research Organization (NARO)
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KUSUMOTO Ken-Ichi
National Food Research Institute, National Agriculture and Food Research Organization (NARO)
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Kusumoto K
National Food Research Institute
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Tada Sawaki
National Food Research Institute
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Suzuki S
National Food Research Institute
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Matsushita Mayumi
National Food Research Institute
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Kuriyama Ken-ichi
Department Of Applied Molecular Biology And Biochemistry Tokyo University Of Agriculture And Technol
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KUSUMOTO Kenichi
National Food Research Institute
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Matsushita Mayumi
National Food Research Institute National Agriculture And Food Research Organization (naro)
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Kusumoto Ken-ichi
National Food Research Institute
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Kashiwagi Y
National Food Research Institute
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Suzuki Satoshi
National Food Research Institute Naro
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Suzuki Satoshi
National Food Research Institute
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