Cargando…

Efficient expression of heterologous genes by the introduction of the endogenous glyceraldehyde-3-phosphate dehydrogenase gene intron 1 in Ganoderma lucidum

BACKGROUND: Ganoderma lucidum, a well-known medicinal mushroom, has received wide attention as a promising cell factory for producing bioactive compounds. However, efficient expression of heterologous genes remains a major challenge in Ganoderma, hindering metabolic regulation research and molecular...

Descripción completa

Detalles Bibliográficos
Autores principales: You, Hao, Sun, Bin, Li, Na, Xu, Jun-Wei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8379801/
https://www.ncbi.nlm.nih.gov/pubmed/34419069
http://dx.doi.org/10.1186/s12934-021-01654-8
_version_ 1783741081420038144
author You, Hao
Sun, Bin
Li, Na
Xu, Jun-Wei
author_facet You, Hao
Sun, Bin
Li, Na
Xu, Jun-Wei
author_sort You, Hao
collection PubMed
description BACKGROUND: Ganoderma lucidum, a well-known medicinal mushroom, has received wide attention as a promising cell factory for producing bioactive compounds. However, efficient expression of heterologous genes remains a major challenge in Ganoderma, hindering metabolic regulation research and molecular breeding of this species. RESULTS: We show that the presence of glyceraldehyde-3-phosphate dehydrogenase gene (gpd) intron 1 at the 5′ end of, the 3′ end of, or within the heterologous phosphinothricin-resistant gene (bar) is efficient for its expression in G. lucidum. The enhanced expression of bar is exhibited by the higher accumulation of mRNA and increased amounts of protein. Moreover, the insertion of the gpd intron 1 in the β-glucuronidase gene (gus) elevates its mRNA accumulation and enzyme activity, which facilitates the use of this reporter gene in Ganoderma. CONCLUSIONS: This study has demonstrated the importance of the introduction of gpd intron 1 for the efficient expression of bar and gus in G. lucidum. The presence of the gpd intron 1 in heterologous genes increases levels of mRNA accumulation and protein expression in basidiomycete Ganoderma. The developed method may be utilized in upregulating the expression of other heterologous genes in Ganoderma. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-021-01654-8.
format Online
Article
Text
id pubmed-8379801
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-83798012021-08-23 Efficient expression of heterologous genes by the introduction of the endogenous glyceraldehyde-3-phosphate dehydrogenase gene intron 1 in Ganoderma lucidum You, Hao Sun, Bin Li, Na Xu, Jun-Wei Microb Cell Fact Research BACKGROUND: Ganoderma lucidum, a well-known medicinal mushroom, has received wide attention as a promising cell factory for producing bioactive compounds. However, efficient expression of heterologous genes remains a major challenge in Ganoderma, hindering metabolic regulation research and molecular breeding of this species. RESULTS: We show that the presence of glyceraldehyde-3-phosphate dehydrogenase gene (gpd) intron 1 at the 5′ end of, the 3′ end of, or within the heterologous phosphinothricin-resistant gene (bar) is efficient for its expression in G. lucidum. The enhanced expression of bar is exhibited by the higher accumulation of mRNA and increased amounts of protein. Moreover, the insertion of the gpd intron 1 in the β-glucuronidase gene (gus) elevates its mRNA accumulation and enzyme activity, which facilitates the use of this reporter gene in Ganoderma. CONCLUSIONS: This study has demonstrated the importance of the introduction of gpd intron 1 for the efficient expression of bar and gus in G. lucidum. The presence of the gpd intron 1 in heterologous genes increases levels of mRNA accumulation and protein expression in basidiomycete Ganoderma. The developed method may be utilized in upregulating the expression of other heterologous genes in Ganoderma. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-021-01654-8. BioMed Central 2021-08-21 /pmc/articles/PMC8379801/ /pubmed/34419069 http://dx.doi.org/10.1186/s12934-021-01654-8 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/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/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://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
You, Hao
Sun, Bin
Li, Na
Xu, Jun-Wei
Efficient expression of heterologous genes by the introduction of the endogenous glyceraldehyde-3-phosphate dehydrogenase gene intron 1 in Ganoderma lucidum
title Efficient expression of heterologous genes by the introduction of the endogenous glyceraldehyde-3-phosphate dehydrogenase gene intron 1 in Ganoderma lucidum
title_full Efficient expression of heterologous genes by the introduction of the endogenous glyceraldehyde-3-phosphate dehydrogenase gene intron 1 in Ganoderma lucidum
title_fullStr Efficient expression of heterologous genes by the introduction of the endogenous glyceraldehyde-3-phosphate dehydrogenase gene intron 1 in Ganoderma lucidum
title_full_unstemmed Efficient expression of heterologous genes by the introduction of the endogenous glyceraldehyde-3-phosphate dehydrogenase gene intron 1 in Ganoderma lucidum
title_short Efficient expression of heterologous genes by the introduction of the endogenous glyceraldehyde-3-phosphate dehydrogenase gene intron 1 in Ganoderma lucidum
title_sort efficient expression of heterologous genes by the introduction of the endogenous glyceraldehyde-3-phosphate dehydrogenase gene intron 1 in ganoderma lucidum
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8379801/
https://www.ncbi.nlm.nih.gov/pubmed/34419069
http://dx.doi.org/10.1186/s12934-021-01654-8
work_keys_str_mv AT youhao efficientexpressionofheterologousgenesbytheintroductionoftheendogenousglyceraldehyde3phosphatedehydrogenasegeneintron1inganodermalucidum
AT sunbin efficientexpressionofheterologousgenesbytheintroductionoftheendogenousglyceraldehyde3phosphatedehydrogenasegeneintron1inganodermalucidum
AT lina efficientexpressionofheterologousgenesbytheintroductionoftheendogenousglyceraldehyde3phosphatedehydrogenasegeneintron1inganodermalucidum
AT xujunwei efficientexpressionofheterologousgenesbytheintroductionoftheendogenousglyceraldehyde3phosphatedehydrogenasegeneintron1inganodermalucidum