A novel plasmid‐based experimental system in Saccharomyces cerevisiae that enables the introduction of 10 different plasmids into cells
The budding yeast Saccharomyces cerevisiae is commonly used as an expression platform for the production of valuable compounds. Yeast‐based genetic research can uniquely utilize auxotrophy in transformant selection: auxotrophic complementation by an auxotrophic marker gene on exogenous DNA (such as...
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| Format: | Article |
| Language: | English |
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Wiley
2024-12-01
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| Series: | FEBS Open Bio |
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| Online Access: | https://doi.org/10.1002/2211-5463.13893 |
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| author | Geyao Dong Tsuyoshi Nakai Tetsuo Matsuzaki |
| author_facet | Geyao Dong Tsuyoshi Nakai Tetsuo Matsuzaki |
| author_sort | Geyao Dong |
| collection | DOAJ |
| description | The budding yeast Saccharomyces cerevisiae is commonly used as an expression platform for the production of valuable compounds. Yeast‐based genetic research can uniquely utilize auxotrophy in transformant selection: auxotrophic complementation by an auxotrophic marker gene on exogenous DNA (such as plasmids). However, the number of required auxotrophic nutrients restricts the number of plasmids maintained by the cells. We, therefore, developed novel Δ10 strains that are auxotrophic for 10 different nutrients and new plasmids with two multiple cloning sites and auxotrophic markers for use in Δ10 strains. We confirmed that Δ10 strains were able to maintain 10 types of plasmids. Using plasmids encoding model proteins, we detected the co‐expression of 17 different genes in Δ10 cell lines. We also constructed Δ9 strains that exhibited auxotrophy for nine nutrients and increased growth compared to Δ10. This study opens a new avenue for the co‐expression of a large number of genes in eukaryotic cells. |
| format | Article |
| id | doaj-art-9f0dac7d14dc46ce981fe664c2f9f359 |
| institution | OA Journals |
| issn | 2211-5463 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Wiley |
| record_format | Article |
| series | FEBS Open Bio |
| spelling | doaj-art-9f0dac7d14dc46ce981fe664c2f9f3592025-08-20T02:18:24ZengWileyFEBS Open Bio2211-54632024-12-0114121955197110.1002/2211-5463.13893A novel plasmid‐based experimental system in Saccharomyces cerevisiae that enables the introduction of 10 different plasmids into cellsGeyao Dong0Tsuyoshi Nakai1Tetsuo Matsuzaki2Department of Neuropsychopharmacology and Hospital Pharmacy Nagoya University Graduate School of Medicine JapanDepartment of Neuropsychopharmacology and Hospital Pharmacy Nagoya University Graduate School of Medicine JapanDepartment of Neuropsychopharmacology and Hospital Pharmacy Nagoya University Graduate School of Medicine JapanThe budding yeast Saccharomyces cerevisiae is commonly used as an expression platform for the production of valuable compounds. Yeast‐based genetic research can uniquely utilize auxotrophy in transformant selection: auxotrophic complementation by an auxotrophic marker gene on exogenous DNA (such as plasmids). However, the number of required auxotrophic nutrients restricts the number of plasmids maintained by the cells. We, therefore, developed novel Δ10 strains that are auxotrophic for 10 different nutrients and new plasmids with two multiple cloning sites and auxotrophic markers for use in Δ10 strains. We confirmed that Δ10 strains were able to maintain 10 types of plasmids. Using plasmids encoding model proteins, we detected the co‐expression of 17 different genes in Δ10 cell lines. We also constructed Δ9 strains that exhibited auxotrophy for nine nutrients and increased growth compared to Δ10. This study opens a new avenue for the co‐expression of a large number of genes in eukaryotic cells.https://doi.org/10.1002/2211-5463.13893auxotrophyplasmidprotein expressionSaccharomyces cerevisiae |
| spellingShingle | Geyao Dong Tsuyoshi Nakai Tetsuo Matsuzaki A novel plasmid‐based experimental system in Saccharomyces cerevisiae that enables the introduction of 10 different plasmids into cells FEBS Open Bio auxotrophy plasmid protein expression Saccharomyces cerevisiae |
| title | A novel plasmid‐based experimental system in Saccharomyces cerevisiae that enables the introduction of 10 different plasmids into cells |
| title_full | A novel plasmid‐based experimental system in Saccharomyces cerevisiae that enables the introduction of 10 different plasmids into cells |
| title_fullStr | A novel plasmid‐based experimental system in Saccharomyces cerevisiae that enables the introduction of 10 different plasmids into cells |
| title_full_unstemmed | A novel plasmid‐based experimental system in Saccharomyces cerevisiae that enables the introduction of 10 different plasmids into cells |
| title_short | A novel plasmid‐based experimental system in Saccharomyces cerevisiae that enables the introduction of 10 different plasmids into cells |
| title_sort | novel plasmid based experimental system in saccharomyces cerevisiae that enables the introduction of 10 different plasmids into cells |
| topic | auxotrophy plasmid protein expression Saccharomyces cerevisiae |
| url | https://doi.org/10.1002/2211-5463.13893 |
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