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Engineering yeast artificial core promoter with designated base motifs
BACKGROUND: Synthetic biology requires toolbox of promoters to finely tune gene expression levels for building up efficient cell factories. Yeast promoters owned variable core promoter regions between the TATA-box and transcriptional starting site (TSS) at the length mostly around 20–80 bases. This...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7026997/ https://www.ncbi.nlm.nih.gov/pubmed/32070349 http://dx.doi.org/10.1186/s12934-020-01305-4 |
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author | Liu, Rui Liu, Lanqing Li, Xia Liu, Duo Yuan, Yingjin |
author_facet | Liu, Rui Liu, Lanqing Li, Xia Liu, Duo Yuan, Yingjin |
author_sort | Liu, Rui |
collection | PubMed |
description | BACKGROUND: Synthetic biology requires toolbox of promoters to finely tune gene expression levels for building up efficient cell factories. Yeast promoters owned variable core promoter regions between the TATA-box and transcriptional starting site (TSS) at the length mostly around 20–80 bases. This region allowed flexible design of artificial promoter but potentially demand special base motifs to maintain or enhance the promoter’s strength. RESULTS: Here, we designed and screened the base motifs and tested the activities of yeast artificial core promoters. Different 30 bases of artificial sequences led to variable expression levels of CrtY enzyme which determined the lycopene–carotene compositions, represented in the colony-color spectrum of red–orange–yellow. The upstream sequences of two strong promoter P(EXP1) and P(GPD) and two starting strains with distinguishable lycopene production levels were utilized to characterize the promoter sequences. Different partition designs of T-rich or G/C-rich base motifs led to distinguishable colony-color distributions. Finally, we screened a champion promoter with a highest 5.5-fold enhancement of lycopene–carotene transformation. Another selected promoter generated a highest beta-carotene production as 7.4 mg/g DCW. CONCLUSIONS: This work offered an approach to redesign promoter with artificial sequences. We concluded that the core promoter region could be designated as 30 bases and different base motifs would enhance or weaken the promoter’s strength. Generally, more T-rich elements, higher %T and lower G/C percentage were beneficial to enhance the strength of artificial core promoter. |
format | Online Article Text |
id | pubmed-7026997 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-70269972020-02-24 Engineering yeast artificial core promoter with designated base motifs Liu, Rui Liu, Lanqing Li, Xia Liu, Duo Yuan, Yingjin Microb Cell Fact Research BACKGROUND: Synthetic biology requires toolbox of promoters to finely tune gene expression levels for building up efficient cell factories. Yeast promoters owned variable core promoter regions between the TATA-box and transcriptional starting site (TSS) at the length mostly around 20–80 bases. This region allowed flexible design of artificial promoter but potentially demand special base motifs to maintain or enhance the promoter’s strength. RESULTS: Here, we designed and screened the base motifs and tested the activities of yeast artificial core promoters. Different 30 bases of artificial sequences led to variable expression levels of CrtY enzyme which determined the lycopene–carotene compositions, represented in the colony-color spectrum of red–orange–yellow. The upstream sequences of two strong promoter P(EXP1) and P(GPD) and two starting strains with distinguishable lycopene production levels were utilized to characterize the promoter sequences. Different partition designs of T-rich or G/C-rich base motifs led to distinguishable colony-color distributions. Finally, we screened a champion promoter with a highest 5.5-fold enhancement of lycopene–carotene transformation. Another selected promoter generated a highest beta-carotene production as 7.4 mg/g DCW. CONCLUSIONS: This work offered an approach to redesign promoter with artificial sequences. We concluded that the core promoter region could be designated as 30 bases and different base motifs would enhance or weaken the promoter’s strength. Generally, more T-rich elements, higher %T and lower G/C percentage were beneficial to enhance the strength of artificial core promoter. BioMed Central 2020-02-18 /pmc/articles/PMC7026997/ /pubmed/32070349 http://dx.doi.org/10.1186/s12934-020-01305-4 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. 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 in a credit line to the data. |
spellingShingle | Research Liu, Rui Liu, Lanqing Li, Xia Liu, Duo Yuan, Yingjin Engineering yeast artificial core promoter with designated base motifs |
title | Engineering yeast artificial core promoter with designated base motifs |
title_full | Engineering yeast artificial core promoter with designated base motifs |
title_fullStr | Engineering yeast artificial core promoter with designated base motifs |
title_full_unstemmed | Engineering yeast artificial core promoter with designated base motifs |
title_short | Engineering yeast artificial core promoter with designated base motifs |
title_sort | engineering yeast artificial core promoter with designated base motifs |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7026997/ https://www.ncbi.nlm.nih.gov/pubmed/32070349 http://dx.doi.org/10.1186/s12934-020-01305-4 |
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