Integrative modules for efficient genome engineering in yeast
We present a set of vectors containing integrative modules for efficient genome integration into the commonly used selection marker loci of the yeast Saccharomyces cerevisiae. A fragment for genome integration is generated via PCR with a unique set of short primers and integrated into HIS3, URA3, AD...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
Shared Science Publishers OG
2017-06-01
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| Series: | Microbial Cell |
| Subjects: | |
| Online Access: | http://microbialcell.com/researcharticles/integrative-modules-for-efficient-genome-engineering-in-yeast/ |
| _version_ | 1811269003014832128 |
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| author | Triana Amen Daniel Kaganovich |
| author_facet | Triana Amen Daniel Kaganovich |
| author_sort | Triana Amen |
| collection | DOAJ |
| description | We present a set of vectors containing integrative modules for efficient genome integration into the commonly used selection marker loci of the yeast Saccharomyces cerevisiae. A fragment for genome integration is generated via PCR with a unique set of short primers and integrated into HIS3, URA3, ADE2, and TRP1 loci. The desired level of expression can be achieved by using constitutive (TEF1p, GPD1p), inducible (CUP1p, GAL1/10p), and daughter-specific (DSE4p) promoters available in the modules. The reduced size of the integrative module compared to conventional integrative plasmids allows efficient integration of multiple fragments. We demonstrate the efficiency of this tool by simultaneously tagging markers of the nucleus, vacuole, actin, and peroxisomes with genomically integrated fluorophores. Improved integration of our new pDK plasmid series allows stable introduction of several genes and can be used for multi-color imaging. New bidirectional promoters (TEF1p-GPD1p, TEF1p-CUP1p, and TEF1p-DSE4p) allow tractable metabolic engineering. |
| first_indexed | 2024-04-12T21:33:14Z |
| format | Article |
| id | doaj.art-1667f9a3b0794c13bfb2ef6b0f7a7801 |
| institution | Directory Open Access Journal |
| issn | 2311-2638 |
| language | English |
| last_indexed | 2024-04-12T21:33:14Z |
| publishDate | 2017-06-01 |
| publisher | Shared Science Publishers OG |
| record_format | Article |
| series | Microbial Cell |
| spelling | doaj.art-1667f9a3b0794c13bfb2ef6b0f7a78012022-12-22T03:15:58ZengShared Science Publishers OGMicrobial Cell2311-26382017-06-014618219010.15698/mic2017.06.576Integrative modules for efficient genome engineering in yeastTriana Amen0Daniel Kaganovich1Department of Cell and Developmental Biology, Hebrew University of Jerusalem, Givat Ram, Jerusalem 91904, Israel.Department of Cell and Developmental Biology, Hebrew University of Jerusalem, Givat Ram, Jerusalem 91904, Israel.We present a set of vectors containing integrative modules for efficient genome integration into the commonly used selection marker loci of the yeast Saccharomyces cerevisiae. A fragment for genome integration is generated via PCR with a unique set of short primers and integrated into HIS3, URA3, ADE2, and TRP1 loci. The desired level of expression can be achieved by using constitutive (TEF1p, GPD1p), inducible (CUP1p, GAL1/10p), and daughter-specific (DSE4p) promoters available in the modules. The reduced size of the integrative module compared to conventional integrative plasmids allows efficient integration of multiple fragments. We demonstrate the efficiency of this tool by simultaneously tagging markers of the nucleus, vacuole, actin, and peroxisomes with genomically integrated fluorophores. Improved integration of our new pDK plasmid series allows stable introduction of several genes and can be used for multi-color imaging. New bidirectional promoters (TEF1p-GPD1p, TEF1p-CUP1p, and TEF1p-DSE4p) allow tractable metabolic engineering.http://microbialcell.com/researcharticles/integrative-modules-for-efficient-genome-engineering-in-yeast/vectorbidirectional promoterintegrative plasmidgenetic integrationyeastSaccharomyces cerevisiae |
| spellingShingle | Triana Amen Daniel Kaganovich Integrative modules for efficient genome engineering in yeast Microbial Cell vector bidirectional promoter integrative plasmid genetic integration yeast Saccharomyces cerevisiae |
| title | Integrative modules for efficient genome engineering in yeast |
| title_full | Integrative modules for efficient genome engineering in yeast |
| title_fullStr | Integrative modules for efficient genome engineering in yeast |
| title_full_unstemmed | Integrative modules for efficient genome engineering in yeast |
| title_short | Integrative modules for efficient genome engineering in yeast |
| title_sort | integrative modules for efficient genome engineering in yeast |
| topic | vector bidirectional promoter integrative plasmid genetic integration yeast Saccharomyces cerevisiae |
| url | http://microbialcell.com/researcharticles/integrative-modules-for-efficient-genome-engineering-in-yeast/ |
| work_keys_str_mv | AT trianaamen integrativemodulesforefficientgenomeengineeringinyeast AT danielkaganovich integrativemodulesforefficientgenomeengineeringinyeast |