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Design and characterization of a synthetic minimal promoter for heterocyst-specific expression in filamentous cyanobacteria
Short and well defined promoters are essential for advancing cyanobacterial biotechnology. The heterocyst of Nostoc sp. is suggested as a microbial cell factory for oxygen sensitive catalysts, such as hydrogenases for hydrogen production, due to its microoxic environment. We identified and predicted...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6133370/ https://www.ncbi.nlm.nih.gov/pubmed/30204806 http://dx.doi.org/10.1371/journal.pone.0203898 |
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author | Wegelius, Adam Li, Xin Turco, Federico Stensjö, Karin |
author_facet | Wegelius, Adam Li, Xin Turco, Federico Stensjö, Karin |
author_sort | Wegelius, Adam |
collection | PubMed |
description | Short and well defined promoters are essential for advancing cyanobacterial biotechnology. The heterocyst of Nostoc sp. is suggested as a microbial cell factory for oxygen sensitive catalysts, such as hydrogenases for hydrogen production, due to its microoxic environment. We identified and predicted promoter elements of possible significance through a consensus strategy using a pool of heterocyst-induced DIF(+) promoters known from Anabaena sp. PCC 7120. To test if these conserved promoter elements were crucial for heterocyst-specific expression, promoter-yfp reporter constructs were designed. The characterization was accomplished by replacing, -35 and -10 regions and the upstream element, with well described elements from the trc promoter of Escherichia coli, which is also functional in Nostoc sp. From the in vivo spatial fluorescence of the different promoter-yfp reporters in Nostoc punctiforme ATCC 29133, we concluded that both the consensus -35 and extended -10 regions were important for heterocyst-specific expression. Further that the promoter strength could be improved by the addition of an upstream element. We designed a short synthetic promoter of 48 nucleotides, P(synDIF), including a consensus DIF1 sequence, a 17 base pair stretch of random nucleotides and an extended consensus -10 region, and thus generated the shortest promoter for heterocyst-specific expression to date. |
format | Online Article Text |
id | pubmed-6133370 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-61333702018-09-27 Design and characterization of a synthetic minimal promoter for heterocyst-specific expression in filamentous cyanobacteria Wegelius, Adam Li, Xin Turco, Federico Stensjö, Karin PLoS One Research Article Short and well defined promoters are essential for advancing cyanobacterial biotechnology. The heterocyst of Nostoc sp. is suggested as a microbial cell factory for oxygen sensitive catalysts, such as hydrogenases for hydrogen production, due to its microoxic environment. We identified and predicted promoter elements of possible significance through a consensus strategy using a pool of heterocyst-induced DIF(+) promoters known from Anabaena sp. PCC 7120. To test if these conserved promoter elements were crucial for heterocyst-specific expression, promoter-yfp reporter constructs were designed. The characterization was accomplished by replacing, -35 and -10 regions and the upstream element, with well described elements from the trc promoter of Escherichia coli, which is also functional in Nostoc sp. From the in vivo spatial fluorescence of the different promoter-yfp reporters in Nostoc punctiforme ATCC 29133, we concluded that both the consensus -35 and extended -10 regions were important for heterocyst-specific expression. Further that the promoter strength could be improved by the addition of an upstream element. We designed a short synthetic promoter of 48 nucleotides, P(synDIF), including a consensus DIF1 sequence, a 17 base pair stretch of random nucleotides and an extended consensus -10 region, and thus generated the shortest promoter for heterocyst-specific expression to date. Public Library of Science 2018-09-11 /pmc/articles/PMC6133370/ /pubmed/30204806 http://dx.doi.org/10.1371/journal.pone.0203898 Text en © 2018 Wegelius et al http://creativecommons.org/licenses/by/4.0/ 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 author and source are credited. |
spellingShingle | Research Article Wegelius, Adam Li, Xin Turco, Federico Stensjö, Karin Design and characterization of a synthetic minimal promoter for heterocyst-specific expression in filamentous cyanobacteria |
title | Design and characterization of a synthetic minimal promoter for heterocyst-specific expression in filamentous cyanobacteria |
title_full | Design and characterization of a synthetic minimal promoter for heterocyst-specific expression in filamentous cyanobacteria |
title_fullStr | Design and characterization of a synthetic minimal promoter for heterocyst-specific expression in filamentous cyanobacteria |
title_full_unstemmed | Design and characterization of a synthetic minimal promoter for heterocyst-specific expression in filamentous cyanobacteria |
title_short | Design and characterization of a synthetic minimal promoter for heterocyst-specific expression in filamentous cyanobacteria |
title_sort | design and characterization of a synthetic minimal promoter for heterocyst-specific expression in filamentous cyanobacteria |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6133370/ https://www.ncbi.nlm.nih.gov/pubmed/30204806 http://dx.doi.org/10.1371/journal.pone.0203898 |
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