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https://hdl.handle.net/20.500.14094/90005177
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2024-04-26
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90005177 (fulltext)
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メタデータID
90005177
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open access
出版タイプ
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タイトル
A novel method for transforming the thermophilic bacterium Geobacillus kaustophilus
著者
Miyano, Megumi ; Tanaka, Kosei ; Ishikawa, Shu ; Mori, Kotaro ; Miguel-Arribas, Andres ; Meijer, Wilfried J. J. ; Yoshida, Ken-ichi
著者名
Miyano, Megumi
著者ID
A1329
研究者ID
1000050418696
著者名
Tanaka, Kosei
田中, 耕生
タナカ, コウセイ
所属機関名
先端融合研究環
著者ID
A0019
研究者ID
1000030359872
KUID
https://kuid-rm-web.ofc.kobe-u.ac.jp/search/detail?systemId=ff864c575b0045a5520e17560c007669
著者名
Ishikawa, Shu
石川, 周
イシカワ, シュウ
所属機関名
科学技術イノベーション研究科
著者名
Mori, Kotaro
著者名
Miguel-Arribas, Andres
著者名
Meijer, Wilfried J. J.
著者ID
A1049
研究者ID
1000020230732
KUID
https://kuid-rm-web.ofc.kobe-u.ac.jp/search/detail?systemId=81199c8c7ea4889b520e17560c007669
著者名
Yoshida, Ken-ichi
吉田, 健一
ヨシダ, ケンイチ
所属機関名
科学技術イノベーション研究科
収録物名
Microbial Cell Factories
巻(号)
17
ページ
127-127
出版者
BMC
刊行日
2018-08-17
公開日
2018-09-05
抄録
Background: Bacterial strains of the genus Geobacillus grow at high temperatures of 50-75 °C and could thus be useful for biotechnological applications. However, genetic manipulation of these species is difficult because the current techniques for transforming Geobacillus species are not efficient. In this study, we developed an easy and efficient method for transforming Geobacillus kaustophilus using the conjugative plasmid pLS20cat. Results: We constructed a transformation system comprising (i) a mobilizable Bacillus subtilis-G. kaustophilus shuttle plasmid named pGK1 that carries the elements for selection and replication in Geobacillus, and (ii) a pLS20cat-har-boring B. subtilis donor strain expressing the dam methylase gene of Escherichia coli and the conjugation-stimulating rap(LS20) gene of pLS20cat. This system can be used to efficiently introduce pGK1 into G. kaustophilus by mobilization in a pLS20cat-dependent way. Whereas the thermostable kanamycin marker and Geobacillus replication origin of pGK1 as well as expression of dam methylase in the donor were indispensable for mobilization, ectopic expression of rap(LS20) increased its efficiency. In addition, the conditions of the recipient influenced mobilization efficiency: the highest mobilization efficiencies were obtained using recipient cells that were in the exponential growth phase. Furthermore, elimination of the origin of transfer from pLS20cat enhanced the mobilization. Conclusions: We describe a novel method of plasmid mobilization into G. kaustophilus recipient from B. subtilis donor depending on the helper function of pLS20cat, which enables simple, rapid, and easy transformation of the thermophilic Gram-positive bacterium.
キーワード
Geobacillus kaustophilus
Bacillus subtilis
Plasmid
Conjugation
Mobilization
Transformation
カテゴリ
科学技術イノベーション研究科
先端融合研究環
学術雑誌論文
権利
© The Author(s) 2018.
This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
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資源タイプ
journal article
言語
English (英語)
eISSN
1475-2859
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関連情報
DOI
https://doi.org/10.1186/s12934-018-0969-9
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