Cloning and molecular characterization of the acetamidase-encoding gene (amdS) from Aspergillus oryzae

Gomi Katsuya, Kitamoto Katsuhiko, Kuinagai Chieko

研究成果: Article査読

41 被引用数 (Scopus)

抄録

We have isolated an acetamidase-encoding gene (amdS) from Aspergillus oryzae by heterologous hybridization using the corresponding Aspergillus nidulans gene as a probe. The gene is located on a 3.5-kb SacI fragment and its nucleotide (nt) sequence was determined. Compared with the A. nidulans amdS gene, the coding region of A. oryzae gene consists of seven exons interrupted by six introns and encodes 545 amino acid (aa) residues. The deduced aa sequence has a high degree of homology with that of the A. nidulans acetamidase protein. Three introns (IVS-1,IVS-2, and IVS-4) exist at the same positions as those of A. nidulans amdS, whilst three additional introns (IVS-3, IVS-5, and IVS-6) are also present. There is no preference in its codon usage (G + C content in the third position of codons is 51%). Gene disruption experiments demonstrate that the resulting mutants show significantly reduced growth on acetamide-containing medium, indicating that the A. oryzae amdS gene encodes a functional acetamidase that is required for acetamide utilization. Transcriptional analysis by Northern blot reveals a 1.8-kb transcript in RNA extracted from mycelium grown in medium containing acetamide or acetate plus β-alanine as the sole carbon and nitrogen sources.

本文言語English
ページ(範囲)91-98
ページ数8
ジャーナルGene
108
1
DOI
出版ステータスPublished - 1991 12 1
外部発表はい

ASJC Scopus subject areas

  • 遺伝学

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