Deletion analysis of the enolase gene (enoA) promoter from the filamentous fungus Aspergillus oryzae

Tomomi Toda, Motoaki Sano, Munechika Honda, Omar Rimoldi, Yong Yang, Midori Yamamoto, Kumiko Takase, Katsuro Hirozumi, Katsuhiko Kitamoto, Toshitaka Minetoki, Katsuya Gomi, Masayuki Machida

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28 Citations (Scopus)

Abstract

The enolase gene (enoA) is one of the most strongly expressed genes in Aspergillus oryzae. To elucidate the transcription regulatory element for this strong expression and the process of glucose induction, the transcription activity of a series of truncated enoA promoters was measured by using the Escherichia coli uidA gene as a reporter. Deletion of a 104-bp region located -224 nt to -121 nt upstream of the translation initiation site caused both a drastic decrease in the β-glucuronidase (GUS) activity and a loss of glucose induction. Northern blot analysis confirmed that the decrease in GUS activity was achieved at the transcriptional level. In addition, electrophoretic gel mobility shift assays indicated that the 104-bp region contained a 15-bp element, to which one or more A. oryzae cellular factors specifically bind. These results suggest that the 15-bp element between -195 nt and -181 nt includes the sequence essential for the transcription regulation of the A. oryzae enoA gene.

Original languageEnglish
Pages (from-to)260-267
Number of pages8
JournalCurrent Genetics
Volume40
Issue number4
DOIs
Publication statusPublished - 2001 Dec 1

Keywords

  • Aspergillus oryzae
  • Electrophoretic gel mobility shift assay (EMSA)
  • Enolase
  • Promoter
  • β-glucuronidase

ASJC Scopus subject areas

  • Genetics

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    Toda, T., Sano, M., Honda, M., Rimoldi, O., Yang, Y., Yamamoto, M., Takase, K., Hirozumi, K., Kitamoto, K., Minetoki, T., Gomi, K., & Machida, M. (2001). Deletion analysis of the enolase gene (enoA) promoter from the filamentous fungus Aspergillus oryzae. Current Genetics, 40(4), 260-267. https://doi.org/10.1007/s00294-001-0258-7