Poulsen, Nicole and Kroeger, Nils (2005) A new molecular tool for transgenic diatoms - Control of mRNA and protein biosynthesis by an inducible promoter-terminator cassette. FEBS JOURNAL, 272 (13). pp. 3413-3423. ISSN 1742-464X, 1742-4658
Full text not available from this repository. (Request a copy)Abstract
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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | CYLINDROTHECA-FUSIFORMIS BACILLARIOPHYCEAE; GREEN FLUORESCENT PROTEIN; NITRATE REDUCTASE GENE; CHLAMYDOMONAS-REINHARDTII; TRANSCRIPTIONAL REGULATION; PHAEODACTYLUM-TRICORNUTUM; THALASSIOSIRA-PSEUDONANA; SACCHAROMYCES-CEREVISIAE; BIOGENIC SILICA; EXPRESSION; Cylindrotheca fusiformis; diatom transformation; green fluorescent protein (GFP); inducible gene expression; nitrate reductase |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie I |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 07 May 2021 06:12 |
| Last Modified: | 07 May 2021 06:12 |
| URI: | https://pred.uni-regensburg.de/id/eprint/35928 |
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