A temperature-regulated replicon-based DNA expression system.
We present a temperature-regulated, alphavirus replicon-based DNA expression system. The system is regulated by a viral temperature-sensitive RNA-dependent RNA replicase, creating a temperature-dependent RNA amplification loop. Because of this positive feedback, the system exhibits both low backgrou...
| Main Authors: | , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2000
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| _version_ | 1797078802113757184 |
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| author | Boorsma, M Nieba, L Koller, D Bachmann, M Bailey, J Renner, W |
| author_facet | Boorsma, M Nieba, L Koller, D Bachmann, M Bailey, J Renner, W |
| author_sort | Boorsma, M |
| collection | OXFORD |
| description | We present a temperature-regulated, alphavirus replicon-based DNA expression system. The system is regulated by a viral temperature-sensitive RNA-dependent RNA replicase, creating a temperature-dependent RNA amplification loop. Because of this positive feedback, the system exhibits both low background and high inducibility. We observed 700-fold induction in transiently transfected cells, and over 104-fold induction in stably transfected cells. The high stringency of inducibility allowed the generation of stable cell lines expressing a highly toxic protein upon temperature shift. These data suggest that the present expression system could simplify bioprocess engineering strategies, especially in situations where the cloned protein has detrimental effects on host cell metabolism. |
| first_indexed | 2024-03-07T00:36:54Z |
| format | Journal article |
| id | oxford-uuid:81bc43bc-45b6-4a17-b3a6-bb50578a5871 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T00:36:54Z |
| publishDate | 2000 |
| record_format | dspace |
| spelling | oxford-uuid:81bc43bc-45b6-4a17-b3a6-bb50578a58712022-03-26T21:32:15ZA temperature-regulated replicon-based DNA expression system.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:81bc43bc-45b6-4a17-b3a6-bb50578a5871EnglishSymplectic Elements at Oxford2000Boorsma, MNieba, LKoller, DBachmann, MBailey, JRenner, WWe present a temperature-regulated, alphavirus replicon-based DNA expression system. The system is regulated by a viral temperature-sensitive RNA-dependent RNA replicase, creating a temperature-dependent RNA amplification loop. Because of this positive feedback, the system exhibits both low background and high inducibility. We observed 700-fold induction in transiently transfected cells, and over 104-fold induction in stably transfected cells. The high stringency of inducibility allowed the generation of stable cell lines expressing a highly toxic protein upon temperature shift. These data suggest that the present expression system could simplify bioprocess engineering strategies, especially in situations where the cloned protein has detrimental effects on host cell metabolism. |
| spellingShingle | Boorsma, M Nieba, L Koller, D Bachmann, M Bailey, J Renner, W A temperature-regulated replicon-based DNA expression system. |
| title | A temperature-regulated replicon-based DNA expression system. |
| title_full | A temperature-regulated replicon-based DNA expression system. |
| title_fullStr | A temperature-regulated replicon-based DNA expression system. |
| title_full_unstemmed | A temperature-regulated replicon-based DNA expression system. |
| title_short | A temperature-regulated replicon-based DNA expression system. |
| title_sort | temperature regulated replicon based dna expression system |
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