Development of a Versatile Expression Plasmid Construction System for<i>Aspergillus oryzae</i>and Its Application to Visualization of Mitochondria

• MABASHI Yuka

  Department of Biotechnology, The University of Tokyo

• KIKUMA Takashi

  Department of Biotechnology, The University of Tokyo

• MARUYAMA Jun-ichi

  Department of Biotechnology, The University of Tokyo

• ARIOKA Manabu

  Department of Biotechnology, The University of Tokyo

• KITAMOTO Katsuhiko

  Department of Biotechnology, The University of Tokyo

書誌事項

タイトル別名

• Development of a Versatile Expression Plasmid Construction System for Aspergillus oryzae and Its Application to Visualization of Mitochondria
• Development of a versatile expression plasmid construction system for <italic>Aspergillus oryzae</italic> and its application to visualization of mitochondria

抄録

We report here a development of the MultiSite Gateway^TM-based versatile plasmid construction system applicable for the rapid and efficient preparation of Aspergillus oryzae expression plasmids. This system allows the simultaneous connection of the three DNA fragments inserted in entry clones along with a destination vector in a defined order and orientation. We prepared a variety of entry clones and destination vectors containing promoters, genes encoding carrier-proteins and fusion tags, and selectable markers, which makes it possible to generate 80 expression plasmids for each target protein. Using this system, plasmids for expression of the EGFP fused with the mitochondrial-targeting signal of citrate synthase (AoCit1) were generated. Tubular structures of mitochondria were visualized in the transformants expressing the AoCit1-EGFP fusion protein. This plasmid construction system allows us to prepare a large number of expression plasmids without laborious DNA manipulations, which would facilitate molecular biological studies on A. oryzae.

収録刊行物

• Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ

  Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ 70 (8), 1882-1889, 2006

  公益社団法人 日本農芸化学会

キーワード

• <I>Aspergillus oryzae</I>
• EGFP
• expression plasmid
• protein expression
• mitochondria

詳細情報

• CRID: 1390001206479497728
• NII論文ID: 10018528059
• NII書誌ID: AA10824164
• DOI: 10.1271/bbb.60052
• COI: 1:CAS:528:DC%2BD28Xpsl2htrc%3D
• ISSN: 13476947
• ISSN: 09168451
• NDL書誌ID: 8040300
• PubMed: 16880596
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I8040300
• Web Site: http://www.tandfonline.com/doi/pdf/10.1271/bbb.60052
• 本文言語コード: en
• データソース種別:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
• 抄録ライセンスフラグ: 使用不可