論文 - 詳細
| RRC ID | 33370 |
|---|---|
| 著者 | Robben J, Massie G, Bosmans E, Wellens B, Volckaert G. |
| タイトル | An Escherichia coli plasmid vector system for high-level production and purification of heterologous peptides fused to active chloramphenicol acetyltransferase. |
| ジャーナル | Gene |
| Abstract |
A very small plasmid vector system is described for construction and high-level production of C-terminal chloramphenicol acetyltransferase (CAT) fusion proteins in Escherichia coli. The only functional elements of the plasmid are a minimal region of the ColE1 origin of DNA replication and the Tn9 cat gene, both under control of a tac promoter. Since C-terminal fusion to CAT does not interfere with chloramphenicol (Cm) resistance, plasmids are maintained under Cm selection. Because of its small size (1392 bp), the system is especially convenient for building and expression of synthetic genes and gene fragments. This concept was utilized to generate a fusion with a synthetic gene encoding the multiple-epitope fragment from the rubella virus E1 membrane protein. Affinity-purified fusion proteins were obtained in mg amounts from 100-ml batches of culture fluid, and incorporated as a specific antigen in a rubella immunoglobulin G enzyme-linked immunosorbent assay. |
| 巻・号 | 126(1) |
| ページ | 109-13 |
| 公開日 | 1993-4-15 |
| DOI | 10.1016/0378-1119(93)90597-v |
| PII | 0378-1119(93)90597-V |
| PMID | 7682530 |
| MeSH | Amino Acid Sequence Base Sequence Chloramphenicol O-Acetyltransferase / genetics* Chloramphenicol O-Acetyltransferase / metabolism Cloning, Molecular DNA, Bacterial / biosynthesis Electrophoresis, Polyacrylamide Gel Epitopes / genetics Escherichia coli / genetics* Genetic Vectors* Isopropyl Thiogalactoside / pharmacology Molecular Sequence Data Plasmids* Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Rubella virus / genetics Viral Envelope Proteins / genetics |
| IF | 2.984 |
| 引用数 | 10 |
| 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 | 3 |
| 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) | pJRtac99 |