Deletion analysis of the promoter of Aspergillus oryzae gene encoding heat shock protein 30
| Autor: | Ken-Ichi Kusumoto, Sawaki Tada, Mayumi Matsushita, Yutaka Kashiwagi, Satoshi Suzuki |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Aspergillus oryzae
TATA box HSP30 Heat-Shock Proteins Molecular Sequence Data CAAT box Bioengineering Biology Applied Microbiology and Biotechnology Fungal Proteins Gene Expression Regulation Fungal Heat shock protein Complementary DNA RNA Messenger Cloning Molecular Heat shock Codon Promoter Regions Genetic Sequence Deletion Binding Sites Base Sequence fungi RNA Fungal Promoter biology.organism_classification Molecular biology CDNA Subtraction Thermodynamics Transcription Factors Biotechnology |
| Zdroj: | Journal of Bioscience and Bioengineering. 107:345-351 |
| ISSN: | 1389-1723 |
| DOI: | 10.1016/j.jbiosc.2008.11.024 |
| Popis: | 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 degrees C) 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 degrees C, but strongly at 40 degrees C. We showed that the promoter of this hsp30 induced heterologous gene expression at high temperatures using beta-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. |
| Databáze: | OpenAIRE |
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