論文 - 詳細
| RRC ID | 33381 |
|---|---|
| 著者 | Kast P. |
| タイトル | pKSS--a second-generation general purpose cloning vector for efficient positive selection of recombinant clones. |
| ジャーナル | Gene |
| Abstract |
A new small plasmid vector (pKSS) for the direct selection of insert-containing plasmid clones is presented. The selection strategy is based on the acquired sensitivity of Escherichia coli cells to p-chloro-phenylalanine (p-Cl-Phe) if they carry a pheS allele encoding a phenylalanyl-tRNA synthetase alpha subunit with relaxed substrate specificity. This pheS allele is present on pKSS. Insertion into, or replacement of, the plasmidial pheS gene by a cloned fragment enables transformed pheS wild-type cells to survive on agar plates containing p-Cl-Phe plus ampicillin. This host strain-independent positive selection of recombinant clones proved to be highly efficient (> 99%) and did not require purification of the vector fragment prior to cloning. The high-copy-number vector pKSS offers a multitude of restriction sites and all of the features for analysis of cloned fragments that stem from the cloning vector pBluescript (Stratagene, La Jolla, CA, USA). Thus, pKSS represents a valuable alternative to previously reported positive-selection vectors; it should prove particularly useful for cloning when expecting a high fraction of cells transformed with non-recombinant vector, and for construction of DNA libraries. |
| 巻・号 | 138(1-2) |
| ページ | 109-14 |
| 公開日 | 1994-1-28 |
| DOI | 10.1016/0378-1119(94)90790-0 |
| PII | 0378-1119(94)90790-0 |
| PMID | 8125286 |
| MeSH | Alleles Base Sequence Cloning, Molecular / methods* Escherichia coli / metabolism* Fenclonine / metabolism Genetic Vectors* Molecular Sequence Data Phenylalanine-tRNA Ligase / biosynthesis Phenylalanine-tRNA Ligase / genetics* Phenylalanine-tRNA Ligase / metabolism Plasmids* Recombinant Fusion Proteins / biosynthesis Recombinant Proteins / biosynthesis* Restriction Mapping Substrate Specificity beta-Galactosidase / biosynthesis |
| IF | 2.984 |
| 引用数 | 70 |
| WOS 分野 | GENETICS & HEREDITY |
| 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 | 3.0 |
| リソース情報 | |
| 原核生物(大腸菌) | pKSS |