Transformation of Aspergillus oryzae through plasmid-mediated complementation of the methonine-auxotrophic mutation

• IIMURA Yuzuru

  National Research Institute of Brewing

• GOMI Katsuya

  National Research Institute of Brewing

• Uzu Hiroshi

  National Research Institute of Brewing

• HARA Shodo

  National Research Institute of Brewing

書誌事項

タイトル別名

• Transformation of Aspergillus oryzae through plasmid-mediated complementation of the methionine-auxotrophic mutation.

抄録

A methionine-auxotrophic mutant (Met^-) of Aspergillus oryzae was transformed to prototrophy (Met⁺) with plasmid pKA5-1 carrying a 3.5Kbp genomic DNA fragment of A. oryzae through treatment of spheroplasts with polyethylene glycol 4000 and CaCl₂. The highest transformation frequency obtained was 32 transformants per μg DNA. We recovered the transforming plasmid from the Met⁺ transformant cell, showing that this plasmid was derived from the original plasmid, pKA5-1. The mitotic stability of the Met⁺ transformant was about 70% after growth without selective pressure; the transforming DNA was lost in the Met^- clone. These findings suggest that the transforming plasmid, pKA5-1, exists in the transformant cell as a self-replicating circular DNA.<br> The gene in the 3.5Kbp DNA fragment, which complements the methionine-auxotrophic mutation, can be used as a selectable marker to construct efficient plasmid vectors for A. oryzae.

収録刊行物

• Agricultural and Biological Chemistry

  Agricultural and Biological Chemistry 51 (2), 323-328, 1987

  公益社団法人 日本農芸化学会

キーワード

• General Agricultural and Biological Sciences
• General Biochemistry, Genetics and Molecular Biology

詳細情報

• CRID: 1390282681439659520
• NII論文ID: 110006323259
• NII書誌ID: AA00515312
• DOI: 10.1271/bbb1961.51.323
• COI: 1:CAS:528:DyaL2sXhslSiur4%3D
• ISSN: 18811280
• ISSN: 00021369
• ISSN: http://id.crossref.org/issn/00021369
• 本文言語コード: en
• データソース種別:
  • JaLC
  • Crossref
  • CiNii Articles
• 抄録ライセンスフラグ: 使用不可