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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 ORCID Kaken ID publons researchmap
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 Kaken ID researchmap
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.
Keywords
Diatom
Genetic transformation
Nitzschia
Multi-pulse electroporation
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.
Published Date
2023-12-10
Publication Title
Marine Biotechnology
Volume
volume25
Issue
issue6
Publisher
Springer Science and Business Media LLC
Start Page
1208
End Page
1219
ISSN
1436-2228
NCID
AA11357643
Content Type
Journal Article
language
English
OAI-PMH Set
岡山大学
Copyright Holders
© The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2023
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isVersionOf https://doi.org/10.1007/s10126-023-10273-w
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
Funder Name
Ministry of Education, Culture, Sports, Science and Technology
Japan Society for the Promotion of Science
Asahi Glass Foundation
助成番号
18K05818