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Coordinated induction of multi-gene pathways in Saccharomyces cerevisiae

Bacterial operons are nature’s tool for regulating and coordinating multi-gene expression in prokaryotes. They are also a gene architecture commonly used in the biosynthesis of many pharmaceutically important compounds and industrially useful chemicals. Despite being an important eukaryotic producti...

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Autores principales: Liang, Jing, Ning, Jonathan C., Zhao, Huimin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3581276/
https://www.ncbi.nlm.nih.gov/pubmed/23262224
http://dx.doi.org/10.1093/nar/gks1293
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author Liang, Jing
Ning, Jonathan C.
Zhao, Huimin
author_facet Liang, Jing
Ning, Jonathan C.
Zhao, Huimin
author_sort Liang, Jing
collection PubMed
description Bacterial operons are nature’s tool for regulating and coordinating multi-gene expression in prokaryotes. They are also a gene architecture commonly used in the biosynthesis of many pharmaceutically important compounds and industrially useful chemicals. Despite being an important eukaryotic production host, Saccharomyces cerevisiae has never had such gene architecture. Here, we report the development of a system to assemble and regulate a multi-gene pathway in S. cerevisiae. Full pathways can be constructed using pre-made parts from a plasmid toolbox. Subsequently, through the use of a yeast strain containing a stably integrated gene switch, the assembled pathway can be regulated using a readily available and inexpensive compound—estradiol—with extremely high sensitivity (10 nM). To demonstrate the use of the system, we assembled the five-gene zeaxanthin biosynthetic pathway in a single step and showed the ligand-dependent coordinated expression of all five genes as well as the tightly regulated production of zeaxanthin. Compared with a previously reported constitutive zeaxanthin pathway, our inducible pathway was shown to have 50-fold higher production level.
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spelling pubmed-35812762013-03-07 Coordinated induction of multi-gene pathways in Saccharomyces cerevisiae Liang, Jing Ning, Jonathan C. Zhao, Huimin Nucleic Acids Res Methods Online Bacterial operons are nature’s tool for regulating and coordinating multi-gene expression in prokaryotes. They are also a gene architecture commonly used in the biosynthesis of many pharmaceutically important compounds and industrially useful chemicals. Despite being an important eukaryotic production host, Saccharomyces cerevisiae has never had such gene architecture. Here, we report the development of a system to assemble and regulate a multi-gene pathway in S. cerevisiae. Full pathways can be constructed using pre-made parts from a plasmid toolbox. Subsequently, through the use of a yeast strain containing a stably integrated gene switch, the assembled pathway can be regulated using a readily available and inexpensive compound—estradiol—with extremely high sensitivity (10 nM). To demonstrate the use of the system, we assembled the five-gene zeaxanthin biosynthetic pathway in a single step and showed the ligand-dependent coordinated expression of all five genes as well as the tightly regulated production of zeaxanthin. Compared with a previously reported constitutive zeaxanthin pathway, our inducible pathway was shown to have 50-fold higher production level. Oxford University Press 2013-02 2012-12-21 /pmc/articles/PMC3581276/ /pubmed/23262224 http://dx.doi.org/10.1093/nar/gks1293 Text en © The Author(s) 2012. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/3.0/), which permits non-commercial reuse, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com.
spellingShingle Methods Online
Liang, Jing
Ning, Jonathan C.
Zhao, Huimin
Coordinated induction of multi-gene pathways in Saccharomyces cerevisiae
title Coordinated induction of multi-gene pathways in Saccharomyces cerevisiae
title_full Coordinated induction of multi-gene pathways in Saccharomyces cerevisiae
title_fullStr Coordinated induction of multi-gene pathways in Saccharomyces cerevisiae
title_full_unstemmed Coordinated induction of multi-gene pathways in Saccharomyces cerevisiae
title_short Coordinated induction of multi-gene pathways in Saccharomyces cerevisiae
title_sort coordinated induction of multi-gene pathways in saccharomyces cerevisiae
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3581276/
https://www.ncbi.nlm.nih.gov/pubmed/23262224
http://dx.doi.org/10.1093/nar/gks1293
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