A set of vectors and strains for chromosomal integration in fission yeast
Abstract The expression of heterologous genes is an important technique in yeast genetics. In fission yeast, the leu1 and ura4 genes have been used mainly as selectable markers for heterologous expression. To expand the repertoire of selection markers available for heterologous expression of genes,...
| প্রধান লেখক: | , , , |
|---|---|
| বিন্যাস: | প্রবন্ধ |
| ভাষা: | English |
| প্রকাশিত: |
Nature Portfolio
2023-06-01
|
| মালা: | Scientific Reports |
| অনলাইন ব্যবহার করুন: | https://doi.org/10.1038/s41598-023-36267-1 |
| _version_ | 1827928741285724160 |
|---|---|
| author | Akihisa Matsuyama Atsushi Hashimoto Shinichi Nishimura Minoru Yoshida |
| author_facet | Akihisa Matsuyama Atsushi Hashimoto Shinichi Nishimura Minoru Yoshida |
| author_sort | Akihisa Matsuyama |
| collection | DOAJ |
| description | Abstract The expression of heterologous genes is an important technique in yeast genetics. In fission yeast, the leu1 and ura4 genes have been used mainly as selectable markers for heterologous expression. To expand the repertoire of selection markers available for heterologous expression of genes, here we developed new host-vector systems employing lys1 and arg3. By employing genome editing with the CRISPR/Cas9 system, we isolated several alleles of lys1 and arg3, each having a critical mutation in the ORF region. In parallel, we developed a set of vectors that complement the amino acid auxotrophy of lys1 and arg3 mutants when integrated into each locus. Using these vectors in combination with the previously developed integration vector pDUAL, we successfully observed the localization of three proteins in a cell simultaneously by fusing them with different fluorescent proteins. Thus, these vectors enable combinatorial expression of heterologous genes, which addresses increasingly diverse experimental challenges. |
| first_indexed | 2024-03-13T06:13:03Z |
| format | Article |
| id | doaj.art-f22e7c1cc36742f094d532a64a136d0c |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-13T06:13:03Z |
| publishDate | 2023-06-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-f22e7c1cc36742f094d532a64a136d0c2023-06-11T11:11:13ZengNature PortfolioScientific Reports2045-23222023-06-011311910.1038/s41598-023-36267-1A set of vectors and strains for chromosomal integration in fission yeastAkihisa Matsuyama0Atsushi Hashimoto1Shinichi Nishimura2Minoru Yoshida3Chemical Genomics Research Group, RIKEN Center for Sustainable Resource ScienceChemical Genomics Research Group, RIKEN Center for Sustainable Resource ScienceLaboratory of Microbiology, Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of TokyoChemical Genomics Research Group, RIKEN Center for Sustainable Resource ScienceAbstract The expression of heterologous genes is an important technique in yeast genetics. In fission yeast, the leu1 and ura4 genes have been used mainly as selectable markers for heterologous expression. To expand the repertoire of selection markers available for heterologous expression of genes, here we developed new host-vector systems employing lys1 and arg3. By employing genome editing with the CRISPR/Cas9 system, we isolated several alleles of lys1 and arg3, each having a critical mutation in the ORF region. In parallel, we developed a set of vectors that complement the amino acid auxotrophy of lys1 and arg3 mutants when integrated into each locus. Using these vectors in combination with the previously developed integration vector pDUAL, we successfully observed the localization of three proteins in a cell simultaneously by fusing them with different fluorescent proteins. Thus, these vectors enable combinatorial expression of heterologous genes, which addresses increasingly diverse experimental challenges.https://doi.org/10.1038/s41598-023-36267-1 |
| spellingShingle | Akihisa Matsuyama Atsushi Hashimoto Shinichi Nishimura Minoru Yoshida A set of vectors and strains for chromosomal integration in fission yeast Scientific Reports |
| title | A set of vectors and strains for chromosomal integration in fission yeast |
| title_full | A set of vectors and strains for chromosomal integration in fission yeast |
| title_fullStr | A set of vectors and strains for chromosomal integration in fission yeast |
| title_full_unstemmed | A set of vectors and strains for chromosomal integration in fission yeast |
| title_short | A set of vectors and strains for chromosomal integration in fission yeast |
| title_sort | set of vectors and strains for chromosomal integration in fission yeast |
| url | https://doi.org/10.1038/s41598-023-36267-1 |
| work_keys_str_mv | AT akihisamatsuyama asetofvectorsandstrainsforchromosomalintegrationinfissionyeast AT atsushihashimoto asetofvectorsandstrainsforchromosomalintegrationinfissionyeast AT shinichinishimura asetofvectorsandstrainsforchromosomalintegrationinfissionyeast AT minoruyoshida asetofvectorsandstrainsforchromosomalintegrationinfissionyeast AT akihisamatsuyama setofvectorsandstrainsforchromosomalintegrationinfissionyeast AT atsushihashimoto setofvectorsandstrainsforchromosomalintegrationinfissionyeast AT shinichinishimura setofvectorsandstrainsforchromosomalintegrationinfissionyeast AT minoruyoshida setofvectorsandstrainsforchromosomalintegrationinfissionyeast |