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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...

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Autores principales: von der Heyde, Eva Laura, Klein, Benjamin, Abram, Lars, Hallmann, Armin
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
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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.
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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
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