論文 - 詳細
| RRC ID | 58474 |
|---|---|
| 著者 | Miyazaki K. |
| タイトル | Molecular engineering of a PheS counterselection marker for improved operating efficiency in Escherichia coli. |
| ジャーナル | Biotechniques |
| Abstract |
Escherichia coli phenylalanyl-tRNA synthetase, a-subunit (ePheS) can be useful as a counterselection marker since its A294G variant misincorporates 4-chloro-phenylalanine (4CP) into cellular proteins during translation, thereby causing cell death. The drawback of this method is that selection must be performed in minimal or semisynthetic medium to avoid interference from phenylalanine in the medium. Here, I reengineered ePheS for improved 4CP incorporation efficiency, obtaining variants (T251A/ A294G and T251S/A294G) that exhibited high lethality in Luria-Bertani medium (LB) containing 4CP. These new variants were superior to the A294G variant when used as a counterselection marker in vector curing experiments. |
| 巻・号 | 58(2) |
| ページ | 86-8 |
| 公開日 | 2015-2-1 |
| DOI | 10.2144/000114257 |
| PII | 000114257 |
| PMID | 25652032 |
| MeSH | Escherichia coli / enzymology* Escherichia coli / genetics* Escherichia coli / growth & development Fluorescence Genes, Bacterial Genetic Engineering / methods* Genetic Markers / genetics Mutant Proteins / genetics Mutant Proteins / metabolism Phenylalanine-tRNA Ligase / genetics* Phenylalanine-tRNA Ligase / metabolism* Selection, Genetic* |
| IF | 1.541 |
| 引用数 | 22 |
| 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 | 9 |
| 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 | |
| 遺伝子材料 | pUC18K_ePAG2 (RDB17947) pUC18Z_ePAG2 (RDB17948) |