Efficient expression of heterologous genes by the introduction of the endogenous glyceraldehyde-3-phosphate dehydrogenase gene intron 1 in Ganoderma lucidum
Abstract 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 m...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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BMC
2021-08-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-021-01654-8 |
| _version_ | 1819124960711409664 |
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| author | Hao You Bin Sun Na Li Jun-Wei Xu |
| author_facet | Hao You Bin Sun Na Li Jun-Wei Xu |
| author_sort | Hao You |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-12-22T07:32:33Z |
| format | Article |
| id | doaj.art-8a238bc4a51a4ab8845522538f300204 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-22T07:32:33Z |
| publishDate | 2021-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-8a238bc4a51a4ab8845522538f3002042022-12-21T18:33:59ZengBMCMicrobial Cell Factories1475-28592021-08-012011810.1186/s12934-021-01654-8Efficient expression of heterologous genes by the introduction of the endogenous glyceraldehyde-3-phosphate dehydrogenase gene intron 1 in Ganoderma lucidumHao You0Bin Sun1Na Li2Jun-Wei Xu3Faculty of Life Science and Technology, Kunming University of Science and TechnologyFaculty of Life Science and Technology, Kunming University of Science and TechnologyFaculty of Science, Kunming University of Science and TechnologyFaculty of Life Science and Technology, Kunming University of Science and TechnologyAbstract 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.https://doi.org/10.1186/s12934-021-01654-8GanodermaExpression of heterologous genesIntronmRNA accumulationProtein expressionEnzyme activity |
| spellingShingle | Hao You Bin Sun Na Li Jun-Wei Xu Efficient expression of heterologous genes by the introduction of the endogenous glyceraldehyde-3-phosphate dehydrogenase gene intron 1 in Ganoderma lucidum Microbial Cell Factories Ganoderma Expression of heterologous genes Intron mRNA accumulation Protein expression Enzyme activity |
| 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 | Ganoderma Expression of heterologous genes Intron mRNA accumulation Protein expression Enzyme activity |
| url | https://doi.org/10.1186/s12934-021-01654-8 |
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