Hybrid Sindbis/Epstein-Barr virus episomal expression vector for inducible production of proteins
Alphavirus vectors are attractive as recombinant protein expression systems due to the high level of gene expression achieved. The combination of two mutations in the viral replicase, which render the replicase noncytopathic and temperature-sensitive, allowed the generation of a DNA-based vector (Cy...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2005-08-01
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| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/05392ST03 |
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| _version_ | 1850153885569646592 |
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| author | Lidia Ivanova Johanna Brändli Philippe Saudan Martin F. Bachmann |
| author_facet | Lidia Ivanova Johanna Brändli Philippe Saudan Martin F. Bachmann |
| author_sort | Lidia Ivanova |
| collection | DOAJ |
| description | Alphavirus vectors are attractive as recombinant protein expression systems due to the high level of gene expression achieved. The combination of two mutations in the viral replicase, which render the replicase noncytopathic and temperature-sensitive, allowed the generation of a DNA-based vector (CytTs) that shows temperature inducible expression. This vector is of significant value for the production of toxic protein. However, like for other stable expression systems, tedious screening of individual cell clones are required in order to get a high producer cell clone. To circumvent this, we generated an episomally replicating vector by introducing an Epstein-Barr virus mini-replicon unit into CytTs. This novel vector allowed rapid generation of cell populations that showed tight regulation of expression and comparable expression levels to the ones achieved with high producer cell clones with CytTs. Moreover, protein production with selected cell populations could easily be scaled-up to a fermentation process. |
| format | Article |
| id | doaj-art-8fd0211efcdb4dfabe5c51dcd59e7d2e |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2005-08-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-8fd0211efcdb4dfabe5c51dcd59e7d2e2025-08-20T02:25:36ZengTaylor & Francis GroupBioTechniques0736-62051940-98182005-08-0139220921210.2144/05392ST03Hybrid Sindbis/Epstein-Barr virus episomal expression vector for inducible production of proteinsLidia Ivanova0Johanna Brändli1Philippe Saudan2Martin F. Bachmann31Cytos Biotechnology AG, Schlieren, Switzerland1Cytos Biotechnology AG, Schlieren, Switzerland1Cytos Biotechnology AG, Schlieren, Switzerland1Cytos Biotechnology AG, Schlieren, SwitzerlandAlphavirus vectors are attractive as recombinant protein expression systems due to the high level of gene expression achieved. The combination of two mutations in the viral replicase, which render the replicase noncytopathic and temperature-sensitive, allowed the generation of a DNA-based vector (CytTs) that shows temperature inducible expression. This vector is of significant value for the production of toxic protein. However, like for other stable expression systems, tedious screening of individual cell clones are required in order to get a high producer cell clone. To circumvent this, we generated an episomally replicating vector by introducing an Epstein-Barr virus mini-replicon unit into CytTs. This novel vector allowed rapid generation of cell populations that showed tight regulation of expression and comparable expression levels to the ones achieved with high producer cell clones with CytTs. Moreover, protein production with selected cell populations could easily be scaled-up to a fermentation process.https://www.future-science.com/doi/10.2144/05392ST03 |
| spellingShingle | Lidia Ivanova Johanna Brändli Philippe Saudan Martin F. Bachmann Hybrid Sindbis/Epstein-Barr virus episomal expression vector for inducible production of proteins BioTechniques |
| title | Hybrid Sindbis/Epstein-Barr virus episomal expression vector for inducible production of proteins |
| title_full | Hybrid Sindbis/Epstein-Barr virus episomal expression vector for inducible production of proteins |
| title_fullStr | Hybrid Sindbis/Epstein-Barr virus episomal expression vector for inducible production of proteins |
| title_full_unstemmed | Hybrid Sindbis/Epstein-Barr virus episomal expression vector for inducible production of proteins |
| title_short | Hybrid Sindbis/Epstein-Barr virus episomal expression vector for inducible production of proteins |
| title_sort | hybrid sindbis epstein barr virus episomal expression vector for inducible production of proteins |
| url | https://www.future-science.com/doi/10.2144/05392ST03 |
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