論文 - 詳細
| RRC ID | 33514 |
|---|---|
| 著者 | Gray MR, Colot HV, Guarente L, Rosbash M. |
| タイトル | Open reading frame cloning: identification, cloning, and expression of open reading frame DNA. |
| ジャーナル | Proc Natl Acad Sci U S A |
| Abstract |
A plasmid was constructed that facilitates the cloning and expression of open reading frame DNA. A DNA fragment containing a bacterial promoter and the amino terminus of the cI gene of bacteriophage lambda was fused to an amino-terminally deleted version of the lacZ gene. An appropriate cloning site was inserted between these two fragments such that a frameshift mutation was introduced upstream of the lacZ-encoding DNA. This cloning vehicle produces a relatively low level of beta-galactosidase activity when introduced into Escherichia coli. The insertion of foreign DNA at the cloning site can reverse the frameshift mutation and generate plasmids that produce a relatively high level of beta-galactosidase activity. A large fraction of these plasmids produce a fusion protein that has a portion of the lambda cI protein at the amino terminus, the foreign protein segment in the middle, and the lacZ polypeptide at the carboxyl terminus. The production of a high level of beta-galactosidase and a large fusion polypeptide guarantees the cloning of a DNA fragment with at least one open reading frame that traverses the entirety of the fragment. Hence, the method can identify, clone, and express (as part of a larger fusion polypeptide) open reading frame DNA from among a large collection of DNA fragments. |
| 巻・号 | 79(21) |
| ページ | 6598-602 |
| 公開日 | 1982-11-1 |
| DOI | 10.1073/pnas.79.21.6598 |
| PMID | 6815653 |
| PMC | PMC347175 |
| MeSH | Base Sequence Cloning, Molecular* Galactosidases / genetics* Genes Genetic Engineering / methods* Lac Operon Mutation Operon beta-Galactosidase / genetics* |
| IF | 9.412 |
| 引用数 | 60 |
| WOS 分野 | BIOLOGY |
| 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 | 26 |
| 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) | pMR100 |