Development of a Versatile Expression Plasmid Construction System for Aspergillus oryzae and Its Application to Visualization of Mitochondria
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- MABASHI Yuka
- Department of Biotechnology, The University of Tokyo
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- KIKUMA Takashi
- Department of Biotechnology, The University of Tokyo
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- MARUYAMA Jun-ichi
- Department of Biotechnology, The University of Tokyo
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- ARIOKA Manabu
- Department of Biotechnology, The University of Tokyo
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- KITAMOTO Katsuhiko
- Department of Biotechnology, The University of Tokyo
Bibliographic Information
- Other Title
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- Development of a Versatile Expression Plasmid Construction System for<i>Aspergillus oryzae</i>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
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Abstract
We report here a development of the MultiSite GatewayTM-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.
Journal
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- Bioscience, Biotechnology, and Biochemistry
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Bioscience, Biotechnology, and Biochemistry 70 (8), 1882-1889, 2006
Japan Society for Bioscience, Biotechnology, and Agrochemistry
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Details 詳細情報について
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- CRID
- 1390001206479497728
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- NII Article ID
- 10018528059
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- NII Book ID
- AA10824164
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- COI
- 1:CAS:528:DC%2BD28Xpsl2htrc%3D
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- ISSN
- 13476947
- 09168451
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- NDL BIB ID
- 8040300
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- PubMed
- 16880596
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
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- Abstract License Flag
- Disallowed