A new molecular tool for transgenic diatoms: Control of mRNA and protein biosynthesis by an inducible promoter–terminator cassette
Open accessAbstract
Research in diatom biology has entered the postgenomic era since the recent completion of the Thalassiosira pseudonana genome project. However, the molecular tools available for genetic manipulation of diatoms are still sparse, impeding the functional analysis of diatom genes in vivo . Here we describe the first method for inducible gene expression in transgenic diatoms. This method uses a DNA cassette containing both promoter ( Pnr ) and terminator ( Tnr ) elements derived from the nitrate reductase gene of the diatom Cylindrotheca fusiformis . By using green fluorescent protein ( gfp ) cDNA as a reporter gene, it is demonstrated that gene expression under the control of the Pnr / Tnr cassette is switched off when cells are grown in the presence of ammonium ions and becomes switched on within 4 h when cells are transferred to medium containing nitrate. Incubating cells in nitrogen‐free medium switches on transcription of the gfp gene, yet gfp mRNA does not become translated into protein. This block on translation is released by the addition of nitrate, resulting in rapid onset of GFP production with a drastically reduced delay time of only 1 h. Altogether we have demonstrated that the Pnr / Tnr cassette enables inducible gene expression and control of both the level and timing of mRNA and protein expression in transgenic diatoms.
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Whole-genome sequences have been released for Thalassiosira pseudonana and Phaeodactylum tricornutum, and stable nuclear transformation has been shown for these two species and a few other diatoms.
“Whole-genome sequences have been released for two species, Thalassiosira pseudonana and Phaeodactylum tricornutum, and recent work has shown stable nuclear transformation of these two species and a few other diatoms [91–94].”
From Biofuels from algae: Challenges and potential (2010) · cites this paper as [93]
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- Authors
- Poulsen, N., & Kröger, N.
- Affiliations
- University of Regensburg (DE)
- Georgia Institute of Technology (US)
- Venue
- The FEBS Journal
- Publisher
- Wiley
- Published
- 2005-06-24
- Volume
- 272
- Issue
- 13
- Pages
- 3413-3423
- ISSN
- 1742-464X (ISSN-L)
- 1742-4658
- OpenAlex
- W1916843334
- Cited by
- 229
- References
- 44
- Citations by year
- 2026: 1 · 2025: 4 · 2024: 6 · 2023: 5 · 2022: 12 · 2021: 8 · 2020: 11 · 2019: 7 · 2018: 13 · 2017: 19 · 2016: 16 · 2015: 24 · 2014: 13 · 2013: 12 · 2012: 10
- FWCI
- 6.32
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- FEBS Journal (publishedVersion)
- PubMed (publishedVersion)
- University of Regensburg Publication Server (University of Regensburg) (acceptedVersion)
- Funders
- Deutsche Forschungsgemeinschaft
- Topics
- Physical Sciences › Materials Science › Biomaterials › Diatoms and Algae Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- inducible gene expression, Cylindrotheca fusiformis, Thalassiosira pseudonana, GFP reporter, ammonium repression, nitrate induction, translational regulation
- MeSH terms
- Amino Acid Sequence, Quaternary Ammonium Compounds, Cloning, Molecular, Molecular Sequence Data, Nitrate Reductases, Nitrates, Promoter Regions, Genetic, Recombinant Fusion Proteins, RNA, Messenger, Terminator Regions, Genetic, Transcription, Genetic, Transformation, Genetic, Protein Biosynthesis, Gene Expression Regulation, Enzymologic, Diatoms, Sequence Homology, Amino Acid, Gene Expression Regulation, Plant, Plants, Genetically Modified, Green Fluorescent Proteins, Nitrate Reductase
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