論文 - 詳細
| RRC ID | 3200 |
|---|---|
| 著者 | Chao YP, Chern JT, Wen CS, Fu H. |
| タイトル | Construction and characterization of thermo-inducible vectors derived from heat-sensitive lacI genes in combination with the T7 A1 promoter. |
| ジャーナル | Biotechnol Bioeng |
| Abstract |
The lack of stringency and the cost of induction are two major disadvantages of using lac-derived vectors for recombinant protein productions. To compensate for these drawbacks, a series of thermo-inducible vectors was developed by coupling heat-sensitive lacI (lacIts) with the T7 A1 promoter on a multiple-copynumber plasmid. The lacIts genes were created by the introduction of Gly187-->Ser substitution along with three alternative mutation sites, Leu233-->Lys, Ala241-->Thr, and Gly265-->Asp, generated by site-directed mutagenesis into the wild-type lacI gene. With the LacZ production as a model, the induction profiles for various vectors containing distinct lacIts exhibited a positive trend as the temperature increased. The fully induced level was achieved by applying the temperature shift from 30 degrees C to 42, 40, or 37 degrees C to the cells harboring the plasmid with the Gly187-->Ser, Ala241-->Thr, or Gly265-->Asp substitution in lacI, respectively. As a result, it produced the maximal LacZ production ranging between 46,000 and 54,000 Miller units, corresponding to a 100- to 400-fold amplification over the uninduced level. As a whole, these novel expression vectors are characterized as having tight regulation and facile inducibility, and their practical usefulness in industrial production of recombinant proteins appears promising. |
| 巻・号 | 79(1) |
| ページ | 1-8 |
| 公開日 | 2002-7-5 |
| DOI | 10.1002/bit.10304 |
| PMID | 17590925 |
| MeSH | Bacterial Proteins / genetics* Bacterial Proteins / metabolism Bacteriophage T7 / genetics* Cloning, Molecular / methods Escherichia coli / genetics Escherichia coli / metabolism* Escherichia coli Proteins Genetic Vectors / genetics Hot Temperature Lac Operon / genetics* Lac Repressors Promoter Regions, Genetic / genetics* Protein Engineering / methods* Recombinant Proteins / biosynthesis Repressor Proteins / genetics* Repressor Proteins / metabolism Transfection / methods beta-Galactosidase / biosynthesis* beta-Galactosidase / genetics |
| IF | 4.002 |
| 引用数 | 17 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 2 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 原核生物(大腸菌) | pHRP308 |