Cargando…
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...
Autores principales: | , , |
---|---|
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 |
_version_ | 1782260399352053760 |
---|---|
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. |
format | Online Article Text |
id | pubmed-3581276 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT liangjing coordinatedinductionofmultigenepathwaysinsaccharomycescerevisiae AT ningjonathanc coordinatedinductionofmultigenepathwaysinsaccharomycescerevisiae AT zhaohuimin coordinatedinductionofmultigenepathwaysinsaccharomycescerevisiae |