論文 - 詳細
| RRC ID | 33426 |
|---|---|
| 著者 | Mujacic M, Cooper KW, Baneyx F. |
| タイトル | Cold-inducible cloning vectors for low-temperature protein expression in Escherichia coli: application to the production of a toxic and proteolytically sensitive fusion protein. |
| ジャーナル | Gene |
| Abstract |
TolAI-beta-lactamase a fusion protein consisting of the inner membrane anchoring domain of the Escherichia coli transenvelope protein TolA followed by TEM-beta-lactamase was found to be toxic and highly unstable when transcribed from the bacteriophage T7 promoter at 37 degrees C. Expression at 15 or 23 degrees C alleviated toxicity, but led to only partial stabilization of the fusion protein. To evaluate the usefulness of cold-shock promoters for the production of proteolytically sensitive proteins at low temperatures, we constructed a set of cloning vectors suitable for rapidly positioning PCR products under cspA transcriptional control. TolAI-beta-lactamase degradation was completely abolished when cspA-driven transcription was induced by temperature downshift to 15 or 23 degrees C. Our results suggest that the cspA promoter system may be a valuable tool for the production of proteins containing membrane-spanning domains or otherwise unstable gene products in E. coli. |
| 巻・号 | 238(2) |
| ページ | 325-32 |
| 公開日 | 1999-10-1 |
| DOI | 10.1016/s0378-1119(99)00328-5 |
| PII | S0378111999003285 |
| PMID | 10570960 |
| MeSH | Bacterial Proteins / genetics* Base Sequence Cloning, Molecular Cold Temperature DNA Primers Escherichia coli / genetics* Escherichia coli Proteins* Genetic Vectors* Hydrolysis Promoter Regions, Genetic Recombinant Fusion Proteins / genetics* Recombinant Fusion Proteins / metabolism Recombinant Fusion Proteins / toxicity Transcription, Genetic / genetics beta-Lactamases / genetics* |
| IF | 2.984 |
| 引用数 | 41 |
| WOS 分野 | GENETICS & HEREDITY |
| リソース情報 | |
| 原核生物(大腸菌) | pCS22 pCS24 |