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Utilization of a Strongly Inducible DDI2 Promoter to Control Gene Expression in Saccharomyces cerevisiae

Regulating target gene expression is a common method in yeast research. In Saccharomyces cerevisiae, there are several widely used regulated expression systems, such as the GAL and Tet-off systems. However, all current expression systems possess some intrinsic deficiencies. We have previously report...

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Autores principales: Lin, Aiyang, Zeng, Chuanwen, Wang, Qian, Zhang, Wenqing, Li, Mengyao, Hanna, Michelle, Xiao, Wei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6250804/
https://www.ncbi.nlm.nih.gov/pubmed/30505295
http://dx.doi.org/10.3389/fmicb.2018.02736
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author Lin, Aiyang
Zeng, Chuanwen
Wang, Qian
Zhang, Wenqing
Li, Mengyao
Hanna, Michelle
Xiao, Wei
author_facet Lin, Aiyang
Zeng, Chuanwen
Wang, Qian
Zhang, Wenqing
Li, Mengyao
Hanna, Michelle
Xiao, Wei
author_sort Lin, Aiyang
collection PubMed
description Regulating target gene expression is a common method in yeast research. In Saccharomyces cerevisiae, there are several widely used regulated expression systems, such as the GAL and Tet-off systems. However, all current expression systems possess some intrinsic deficiencies. We have previously reported that the DDI2 gene can be induced to very high levels upon cyanamide or methyl methanesulfonate treatment. Here we report the construction of gene expression systems based on the DDI2 promoter in both single- and multi-copy plasmids. Using GFP as a reporter gene, it was demonstrated that the target gene expression could be increased by up to 2,000-fold at the transcriptional level by utilizing the above systems. In addition, a DDI2-based construct was created for promoter shuffling in the budding yeast genome to control endogenous gene expression. Overall, this study offers a set of convenient and highly efficient experimental tools to control target gene expression in budding yeast.
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spelling pubmed-62508042018-11-30 Utilization of a Strongly Inducible DDI2 Promoter to Control Gene Expression in Saccharomyces cerevisiae Lin, Aiyang Zeng, Chuanwen Wang, Qian Zhang, Wenqing Li, Mengyao Hanna, Michelle Xiao, Wei Front Microbiol Microbiology Regulating target gene expression is a common method in yeast research. In Saccharomyces cerevisiae, there are several widely used regulated expression systems, such as the GAL and Tet-off systems. However, all current expression systems possess some intrinsic deficiencies. We have previously reported that the DDI2 gene can be induced to very high levels upon cyanamide or methyl methanesulfonate treatment. Here we report the construction of gene expression systems based on the DDI2 promoter in both single- and multi-copy plasmids. Using GFP as a reporter gene, it was demonstrated that the target gene expression could be increased by up to 2,000-fold at the transcriptional level by utilizing the above systems. In addition, a DDI2-based construct was created for promoter shuffling in the budding yeast genome to control endogenous gene expression. Overall, this study offers a set of convenient and highly efficient experimental tools to control target gene expression in budding yeast. Frontiers Media S.A. 2018-11-16 /pmc/articles/PMC6250804/ /pubmed/30505295 http://dx.doi.org/10.3389/fmicb.2018.02736 Text en Copyright © 2018 Lin, Zeng, Wang, Zhang, Li, Hanna and Xiao. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Lin, Aiyang
Zeng, Chuanwen
Wang, Qian
Zhang, Wenqing
Li, Mengyao
Hanna, Michelle
Xiao, Wei
Utilization of a Strongly Inducible DDI2 Promoter to Control Gene Expression in Saccharomyces cerevisiae
title Utilization of a Strongly Inducible DDI2 Promoter to Control Gene Expression in Saccharomyces cerevisiae
title_full Utilization of a Strongly Inducible DDI2 Promoter to Control Gene Expression in Saccharomyces cerevisiae
title_fullStr Utilization of a Strongly Inducible DDI2 Promoter to Control Gene Expression in Saccharomyces cerevisiae
title_full_unstemmed Utilization of a Strongly Inducible DDI2 Promoter to Control Gene Expression in Saccharomyces cerevisiae
title_short Utilization of a Strongly Inducible DDI2 Promoter to Control Gene Expression in Saccharomyces cerevisiae
title_sort utilization of a strongly inducible ddi2 promoter to control gene expression in saccharomyces cerevisiae
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6250804/
https://www.ncbi.nlm.nih.gov/pubmed/30505295
http://dx.doi.org/10.3389/fmicb.2018.02736
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