ID | 66234 |
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Okada, Koki
Graduate School of Environmental and Life Science, Okayama University
Morimoto, Yu
Graduate School of Environmental and Life Science, Okayama University
Shiraishi, Yukine
Graduate School of Environmental and Life Science, Okayama University
Tamura, Takashi
Faculty of Environmental, Life, Natural Science and Technology, Okayama University
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Mayama, Shigeki
The Advanced Support Center for Science Teachers, Tokyo Gakugei University
Kadono, Takashi
Faculty of Agriculture and Marine Science, Kochi University
Adachi, Masao
Faculty of Agriculture and Marine Science, Kochi University
Ifuku, Kentaro
Graduate School of Agriculture, Kyoto University
Nemoto, Michiko
Faculty of Environmental, Life, Natural Science and Technology, Okayama University
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Abstract | Nitzschia is one of the largest genera of diatoms found in a range of aquatic environments, from freshwater to seawater. This genus contains evolutionarily and ecologically unique species, such as those that have lost photosynthetic capacity or those that live symbiotically in dinoflagellates. Several Nitzschia species have been used as indicators of water pollution. Recently, Nitzschia species have attracted considerable attention in the field of biotechnology. In this study, a transformation method for the marine pennate diatom Nitzschia sp. strain NIES-4635, isolated from the coastal Seto Inland Sea, was established. Plasmids containing the promoter/terminator of the fucoxanthin chlorophyll a/c binding protein gene (fcp, or Lhcf) derived from Nitzschia palea were constructed and introduced into cells by multi-pulse electroporation, resulting in 500 μg/mL nourseothricin-resistant transformants with transformation frequencies of up to 365 colonies per 108 cells. In addition, when transformation was performed using a new plasmid containing a promoter derived from a diatom-infecting virus upstream of the green fluorescent protein gene (gfp), 44% of the nourseothricin-resistant clones exhibited GFP fluorescence. The integration of the genes introduced into the genomes of the transformants was confirmed by Southern blotting. The Nitzschia transformation method established in this study will enable the transformation this species, thus allowing the functional analysis of genes from the genus Nitzschia, which are important species for environmental and biotechnological development.
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Keywords | Diatom
Genetic transformation
Nitzschia
Multi-pulse electroporation
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Note | This version of the article has been accepted for publication, after peer review (when applicable) and is subject to Springer Nature’s AM terms of use, but is not the Version of Record and does not reflect post-acceptance improvements, or any corrections. The Version of Record is available online at: http://dx.doi.org/10.1007/s10126-023-10273-w
This fulltext file will be available in Dec. 2024.
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Published Date | 2023-12-10
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Publication Title |
Marine Biotechnology
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Volume | volume25
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Issue | issue6
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Publisher | Springer Science and Business Media LLC
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Start Page | 1208
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End Page | 1219
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ISSN | 1436-2228
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NCID | AA11357643
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Content Type |
Journal Article
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language |
English
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OAI-PMH Set |
岡山大学
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Copyright Holders | © The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2023
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File Version | author
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Related Url | isVersionOf https://doi.org/10.1007/s10126-023-10273-w
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Citation | Okada, K., Morimoto, Y., Shiraishi, Y. et al. Nuclear Transformation of the Marine Pennate Diatom Nitzschia sp. Strain NIES-4635 by Multi-Pulse Electroporation. Mar Biotechnol 25, 1208–1219 (2023). https://doi.org/10.1007/s10126-023-10273-w
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Funder Name |
Ministry of Education, Culture, Sports, Science and Technology
Japan Society for the Promotion of Science
Asahi Glass Foundation
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助成番号 | 18K05818
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