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Changes to the core and flanking sequences of G‐box elements lead to increases and decreases in gene expression in both native and synthetic soybean promoters

Cis‐regulatory elements in promoters are major determinants of binding specificity of transcription factors (TFs) for transcriptional regulation. To improve our understanding of how these short DNA sequences regulate gene expression, synthetic promoters consisting of both classical (CACGTG) and vari...

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Autores principales: Zhang, Ning, McHale, Leah K., Finer, John J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6419578/
https://www.ncbi.nlm.nih.gov/pubmed/30191675
http://dx.doi.org/10.1111/pbi.13010
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author Zhang, Ning
McHale, Leah K.
Finer, John J.
author_facet Zhang, Ning
McHale, Leah K.
Finer, John J.
author_sort Zhang, Ning
collection PubMed
description Cis‐regulatory elements in promoters are major determinants of binding specificity of transcription factors (TFs) for transcriptional regulation. To improve our understanding of how these short DNA sequences regulate gene expression, synthetic promoters consisting of both classical (CACGTG) and variant G‐box core sequences along with different flanking sequences derived from the promoters of three different highly expressing soybean genes, were constructed and used to regulate a green fluorescent protein (gfp) gene. Use of the classical 6‐bp G‐box provided information on the base level of GFP expression while modifications to the 2–4 flanking bases on either side of the G‐box influenced the intensity of gene expression in both transiently transformed lima bean cotyledons and stably transformed soybean hairy roots. The proximal 2‐bp sequences on either flank of the G‐box significantly affected G‐box activity, while the distal 2‐bp flanking nucleotides also influenced gene expression albeit with a decreasing effect. Manipulation of the upstream 2‐ to 4‐bp flanking sequence of a G‐box variant (GACGTG), found in the proximal region of a relatively weak soybean glycinin promoter, significantly enhanced promoter activity using both transient and stable expression assays, if the G‐box variant was first converted into a classical G‐box (CACGTG). In addition to increasing our understanding of regulatory element composition and structure, this study shows that minimal targeted changes in native promoter sequences can lead to enhanced gene expression, and suggests that genome editing of the promoter region can result in useful and predictable changes in native gene expression.
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spelling pubmed-64195782019-03-18 Changes to the core and flanking sequences of G‐box elements lead to increases and decreases in gene expression in both native and synthetic soybean promoters Zhang, Ning McHale, Leah K. Finer, John J. Plant Biotechnol J Research Articles Cis‐regulatory elements in promoters are major determinants of binding specificity of transcription factors (TFs) for transcriptional regulation. To improve our understanding of how these short DNA sequences regulate gene expression, synthetic promoters consisting of both classical (CACGTG) and variant G‐box core sequences along with different flanking sequences derived from the promoters of three different highly expressing soybean genes, were constructed and used to regulate a green fluorescent protein (gfp) gene. Use of the classical 6‐bp G‐box provided information on the base level of GFP expression while modifications to the 2–4 flanking bases on either side of the G‐box influenced the intensity of gene expression in both transiently transformed lima bean cotyledons and stably transformed soybean hairy roots. The proximal 2‐bp sequences on either flank of the G‐box significantly affected G‐box activity, while the distal 2‐bp flanking nucleotides also influenced gene expression albeit with a decreasing effect. Manipulation of the upstream 2‐ to 4‐bp flanking sequence of a G‐box variant (GACGTG), found in the proximal region of a relatively weak soybean glycinin promoter, significantly enhanced promoter activity using both transient and stable expression assays, if the G‐box variant was first converted into a classical G‐box (CACGTG). In addition to increasing our understanding of regulatory element composition and structure, this study shows that minimal targeted changes in native promoter sequences can lead to enhanced gene expression, and suggests that genome editing of the promoter region can result in useful and predictable changes in native gene expression. John Wiley and Sons Inc. 2018-09-24 2019-04 /pmc/articles/PMC6419578/ /pubmed/30191675 http://dx.doi.org/10.1111/pbi.13010 Text en © 2018 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Zhang, Ning
McHale, Leah K.
Finer, John J.
Changes to the core and flanking sequences of G‐box elements lead to increases and decreases in gene expression in both native and synthetic soybean promoters
title Changes to the core and flanking sequences of G‐box elements lead to increases and decreases in gene expression in both native and synthetic soybean promoters
title_full Changes to the core and flanking sequences of G‐box elements lead to increases and decreases in gene expression in both native and synthetic soybean promoters
title_fullStr Changes to the core and flanking sequences of G‐box elements lead to increases and decreases in gene expression in both native and synthetic soybean promoters
title_full_unstemmed Changes to the core and flanking sequences of G‐box elements lead to increases and decreases in gene expression in both native and synthetic soybean promoters
title_short Changes to the core and flanking sequences of G‐box elements lead to increases and decreases in gene expression in both native and synthetic soybean promoters
title_sort changes to the core and flanking sequences of g‐box elements lead to increases and decreases in gene expression in both native and synthetic soybean promoters
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6419578/
https://www.ncbi.nlm.nih.gov/pubmed/30191675
http://dx.doi.org/10.1111/pbi.13010
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