論文 - 詳細
| RRC ID | 33353 |
|---|---|
| 著者 | Dersch P, Fsihi H, Bremer E. |
| タイトル | Low-copy-number T7 vectors for selective gene expression and efficient protein overproduction in Escherichia coli. |
| ジャーナル | FEMS Microbiol Lett |
| Abstract |
A set of low-copy-number vectors (pPD) has been constructed that permit selective gene expression and high-level protein overproduction in Escherichia coli, based on the bacteriophage T7 RNA polymerase/T7 promoter system. These plasmids carry a chloramphenicol resistance gene (cat) as a selective marker and an extended multiple cloning site for convenient gene cloning. Their replication is mediated by ori sequences derived from the low-copy-number vector pSC101. The efficient T7 gene 10 promoter present on these vectors allows selective and high-level transcription of cloned genes carrying their own translational initiation signals. In addition, low-copy-number T7 vectors were constructed that permit expression of genes lacking their own transcription and translation initiation elements by providing a ribosome binding site, an ATG start codon and a multiple cloning site devised for the cloning in all three reading frames. The pPD expression vectors were used to achieve high-level overproduction of the E. coli integral outer membrane protein Tsx, and the cytoplasmic enzymes beta-galactosidase (beta Gal) and UTP:alpha-D-glucose-1-phosphate uridylyltransferase (GalU). The characteristics of these low-copy-number T7 expression vectors should prove very useful for the cloning and high-level overexpression of genes whose gene products are deleterious to the E. coli host. |
| 巻・号 | 123(1-2) |
| ページ | 19-26 |
| 公開日 | 1994-10-15 |
| DOI | 10.1111/j.1574-6968.1994.tb07195.x |
| PMID | 7988888 |
| MeSH | Bacterial Outer Membrane Proteins / biosynthesis Bacterial Outer Membrane Proteins / genetics Bacterial Proteins / biosynthesis* Bacterial Proteins / genetics Bacteriophage T7 / genetics* Base Sequence Escherichia coli / genetics* Escherichia coli / metabolism Escherichia coli Proteins* Gene Expression Regulation, Bacterial Molecular Sequence Data Plasmids / genetics Receptors, Virus* beta-Galactosidase / biosynthesis beta-Galactosidase / genetics |
| IF | 1.987 |
| 引用数 | 30 |
| WOS 分野 | MICROBIOLOGY |
| リソース情報 | |
| 原核生物(大腸菌) | pPD1 pPD100 pPD101 pPD110 pPD111 pPD112 |