Cargando…
The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri
BACKGROUND: The multicellular green alga Volvox carteri represents an attractive model system to study various aspects of multicellularity like cellular differentiation, morphogenesis, epithelial folding and ECM biogenesis. However, functional and molecular analyses of such processes require a wide...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4339647/ https://www.ncbi.nlm.nih.gov/pubmed/25888095 http://dx.doi.org/10.1186/s12896-015-0122-3 |
_version_ | 1782358895848587264 |
---|---|
author | von der Heyde, Eva Laura Klein, Benjamin Abram, Lars Hallmann, Armin |
author_facet | von der Heyde, Eva Laura Klein, Benjamin Abram, Lars Hallmann, Armin |
author_sort | von der Heyde, Eva Laura |
collection | PubMed |
description | BACKGROUND: The multicellular green alga Volvox carteri represents an attractive model system to study various aspects of multicellularity like cellular differentiation, morphogenesis, epithelial folding and ECM biogenesis. However, functional and molecular analyses of such processes require a wide array of molecular tools for genetic engineering. So far there are only a limited number of molecular tools available in Volvox. RESULTS: Here, we show that the promoter of the V. carteri nitrate reductase gene (nitA) is a powerful molecular switch for induction of transgene expression. Strong expression is triggered by simply changing the nitrogen source from ammonium to nitrate. We also show that the luciferase (g-luc) gene from the marine copepod Gaussia princeps, which previously was engineered to match the codon usage of the unicellular alga Chlamydomonas reinhardtii, is a suitable reporter gene in V. carteri. Emitted light of the chemiluminescent reaction can be easily detected and quantified with a luminometer. Long-term stability of inducible expression of the chimeric nitA/g-luc transgenes after stable nuclear transformation was demonstrated by transcription analysis and bioluminescence assays. CONCLUSION: Two novel molecular tools for genetic engineering of Volvox are now available: the nitrate-inducible nitA promoter of V. carteri and the codon-adapted luciferase reporter gene of G. princeps. These novel tools will be useful for future molecular research in V. carteri. |
format | Online Article Text |
id | pubmed-4339647 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-43396472015-02-26 The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri von der Heyde, Eva Laura Klein, Benjamin Abram, Lars Hallmann, Armin BMC Biotechnol Methodology Article BACKGROUND: The multicellular green alga Volvox carteri represents an attractive model system to study various aspects of multicellularity like cellular differentiation, morphogenesis, epithelial folding and ECM biogenesis. However, functional and molecular analyses of such processes require a wide array of molecular tools for genetic engineering. So far there are only a limited number of molecular tools available in Volvox. RESULTS: Here, we show that the promoter of the V. carteri nitrate reductase gene (nitA) is a powerful molecular switch for induction of transgene expression. Strong expression is triggered by simply changing the nitrogen source from ammonium to nitrate. We also show that the luciferase (g-luc) gene from the marine copepod Gaussia princeps, which previously was engineered to match the codon usage of the unicellular alga Chlamydomonas reinhardtii, is a suitable reporter gene in V. carteri. Emitted light of the chemiluminescent reaction can be easily detected and quantified with a luminometer. Long-term stability of inducible expression of the chimeric nitA/g-luc transgenes after stable nuclear transformation was demonstrated by transcription analysis and bioluminescence assays. CONCLUSION: Two novel molecular tools for genetic engineering of Volvox are now available: the nitrate-inducible nitA promoter of V. carteri and the codon-adapted luciferase reporter gene of G. princeps. These novel tools will be useful for future molecular research in V. carteri. BioMed Central 2015-02-18 /pmc/articles/PMC4339647/ /pubmed/25888095 http://dx.doi.org/10.1186/s12896-015-0122-3 Text en © von der Heyde et al.; licensee BioMed Central. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. 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. |
spellingShingle | Methodology Article von der Heyde, Eva Laura Klein, Benjamin Abram, Lars Hallmann, Armin The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri |
title | The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri |
title_full | The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri |
title_fullStr | The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri |
title_full_unstemmed | The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri |
title_short | The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri |
title_sort | inducible nita promoter provides a powerful molecular switch for transgene expression in volvox carteri |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4339647/ https://www.ncbi.nlm.nih.gov/pubmed/25888095 http://dx.doi.org/10.1186/s12896-015-0122-3 |
work_keys_str_mv | AT vonderheydeevalaura theinduciblenitapromoterprovidesapowerfulmolecularswitchfortransgeneexpressioninvolvoxcarteri AT kleinbenjamin theinduciblenitapromoterprovidesapowerfulmolecularswitchfortransgeneexpressioninvolvoxcarteri AT abramlars theinduciblenitapromoterprovidesapowerfulmolecularswitchfortransgeneexpressioninvolvoxcarteri AT hallmannarmin theinduciblenitapromoterprovidesapowerfulmolecularswitchfortransgeneexpressioninvolvoxcarteri AT vonderheydeevalaura induciblenitapromoterprovidesapowerfulmolecularswitchfortransgeneexpressioninvolvoxcarteri AT kleinbenjamin induciblenitapromoterprovidesapowerfulmolecularswitchfortransgeneexpressioninvolvoxcarteri AT abramlars induciblenitapromoterprovidesapowerfulmolecularswitchfortransgeneexpressioninvolvoxcarteri AT hallmannarmin induciblenitapromoterprovidesapowerfulmolecularswitchfortransgeneexpressioninvolvoxcarteri |