Transformation of diatom Phaeodactylum tricornutum by electroporation and establishment of inducible selection marker
Diatoms are important primary producers in the marine ecosystem. Currently it is difficult to genetically transform diatoms due to the technical limitations of existing methods. The promoter/terminator of the nitrate reductase gene of the model diatom Phaeodactylum tricornutum was cloned and used to...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2012-06-01
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| Series: | BioTechniques |
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| Online Access: | https://www.future-science.com/doi/10.2144/000113881 |
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| author | Ying-Fang Niu Zhi-Kai Yang Meng-Han Zhang Cong-Cong Zhu Wei-Dong Yang Jie-Sheng Liu Hong-Ye Li |
| author_facet | Ying-Fang Niu Zhi-Kai Yang Meng-Han Zhang Cong-Cong Zhu Wei-Dong Yang Jie-Sheng Liu Hong-Ye Li |
| author_sort | Ying-Fang Niu |
| collection | DOAJ |
| description | Diatoms are important primary producers in the marine ecosystem. Currently it is difficult to genetically transform diatoms due to the technical limitations of existing methods. The promoter/terminator of the nitrate reductase gene of the model diatom Phaeodactylum tricornutum was cloned and used to drive chloramphenicol acetyltransferase (CAT) reporter gene expression. The construct was transferred by electroporation into P. tricornutum grown in medium lacking silicon. CAT expression was induced in transformed diatoms in the presence of nitrate, enabling growth in selective medium, and was repressed when ammonium was the only nitrogen source. Expression of CAT transcript and protein were demonstrated by RT-PCR and Western blot analysis, respectively. Our study is the first to report a successful genetic transformation of diatom by electroporation in an economical and efficient manner and provides a tightly regulated inducible gene expression system for diatom. |
| format | Article |
| id | doaj-art-e11acce75165464fada8ba89225d8907 |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2012-06-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-e11acce75165464fada8ba89225d89072025-08-20T02:25:55ZengTaylor & Francis GroupBioTechniques0736-62051940-98182012-06-015261310.2144/000113881Transformation of diatom Phaeodactylum tricornutum by electroporation and establishment of inducible selection markerYing-Fang Niu0Zhi-Kai Yang1Meng-Han Zhang2Cong-Cong Zhu3Wei-Dong Yang4Jie-Sheng Liu5Hong-Ye Li61Key Laboratory of Eutrophication and Control of HAB of Guangdong Higher Education Institute, Jinan University, Guangzhou, China1Key Laboratory of Eutrophication and Control of HAB of Guangdong Higher Education Institute, Jinan University, Guangzhou, China1Key Laboratory of Eutrophication and Control of HAB of Guangdong Higher Education Institute, Jinan University, Guangzhou, China1Key Laboratory of Eutrophication and Control of HAB of Guangdong Higher Education Institute, Jinan University, Guangzhou, China1Key Laboratory of Eutrophication and Control of HAB of Guangdong Higher Education Institute, Jinan University, Guangzhou, China1Key Laboratory of Eutrophication and Control of HAB of Guangdong Higher Education Institute, Jinan University, Guangzhou, China1Key Laboratory of Eutrophication and Control of HAB of Guangdong Higher Education Institute, Jinan University, Guangzhou, ChinaDiatoms are important primary producers in the marine ecosystem. Currently it is difficult to genetically transform diatoms due to the technical limitations of existing methods. The promoter/terminator of the nitrate reductase gene of the model diatom Phaeodactylum tricornutum was cloned and used to drive chloramphenicol acetyltransferase (CAT) reporter gene expression. The construct was transferred by electroporation into P. tricornutum grown in medium lacking silicon. CAT expression was induced in transformed diatoms in the presence of nitrate, enabling growth in selective medium, and was repressed when ammonium was the only nitrogen source. Expression of CAT transcript and protein were demonstrated by RT-PCR and Western blot analysis, respectively. Our study is the first to report a successful genetic transformation of diatom by electroporation in an economical and efficient manner and provides a tightly regulated inducible gene expression system for diatom.https://www.future-science.com/doi/10.2144/000113881diatomelectroporationnitrate reductase promoterinducible expression |
| spellingShingle | Ying-Fang Niu Zhi-Kai Yang Meng-Han Zhang Cong-Cong Zhu Wei-Dong Yang Jie-Sheng Liu Hong-Ye Li Transformation of diatom Phaeodactylum tricornutum by electroporation and establishment of inducible selection marker BioTechniques diatom electroporation nitrate reductase promoter inducible expression |
| title | Transformation of diatom Phaeodactylum tricornutum by electroporation and establishment of inducible selection marker |
| title_full | Transformation of diatom Phaeodactylum tricornutum by electroporation and establishment of inducible selection marker |
| title_fullStr | Transformation of diatom Phaeodactylum tricornutum by electroporation and establishment of inducible selection marker |
| title_full_unstemmed | Transformation of diatom Phaeodactylum tricornutum by electroporation and establishment of inducible selection marker |
| title_short | Transformation of diatom Phaeodactylum tricornutum by electroporation and establishment of inducible selection marker |
| title_sort | transformation of diatom phaeodactylum tricornutum by electroporation and establishment of inducible selection marker |
| topic | diatom electroporation nitrate reductase promoter inducible expression |
| url | https://www.future-science.com/doi/10.2144/000113881 |
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