論文 - 詳細
| RRC ID | 33424 |
|---|---|
| 著者 | Cote-Sierra J, Jongert E, Bredan A, Gautam DC, Parkhouse M, Cornelis P, De Baetselier P, Revets H. |
| タイトル | A new membrane-bound OprI lipoprotein expression vector. High production of heterologous fusion proteins in gram (-) bacteria and the implications for oral vaccination. |
| ジャーナル | Gene |
| Abstract |
We have previously described the development of cloning vectors for the production of OprI-based outer membrane fusion proteins in E. coli (Cornelis et al., 1996) and now describe the construction of a new vector, containing a lacI(q) gene, resulting in tight repression of the promotor and allowing its use in other Gram (-) bacteria. The new pVUB3 expression vector encodes a truncated but active LacI(q)(341) repressor which binds to the single operator in the vector. A high repression of the trc promotor was observed, resulting in a very low basal leakage of expression and very high production levels of OprI or derivatives after IPTG induction in E. coli. Bacterial viability was not affected under uninduced conditions, but the number of viable cell counts decreased after production of large amounts of the outer membrane-bound OprI lipoprotein and its derivatives, both in E. coli and Salmonella typhimurium. This highly repressible system allows us to extend the use of OprI vectors in other Gram (-) bacteria, resulting in the production of outer membrane-bound lipid-modified molecules, opening the possibility for its application in the design of potential live Salmonella-based subunit vaccines. |
| 巻・号 | 221(1) |
| ページ | 25-34 |
| 公開日 | 1998-10-9 |
| DOI | 10.1016/s0378-1119(98)00437-5 |
| PII | S0378-1119(98)00437-5 |
| PMID | 9852946 |
| MeSH | Amino Acid Sequence Animals Antigens, Protozoan / genetics Antigens, Surface / genetics Bacterial Proteins / chemistry Bacterial Proteins / genetics* Base Sequence Biological Transport Capsid / genetics Capsid / immunology Capsid Proteins* Cell Division / drug effects Cloning, Molecular Escherichia coli / cytology Escherichia coli / drug effects Escherichia coli / genetics Escherichia coli Proteins* Gene Expression Regulation, Bacterial / drug effects Genetic Vectors / drug effects Genetic Vectors / genetics* Genetic Vectors / immunology Gram-Negative Bacteria / genetics* Isopropyl Thiogalactoside / pharmacology Lac Repressors Leishmania major / genetics Lipoproteins / genetics* Molecular Sequence Data Peptide Fragments / genetics Peptide Fragments / immunology Plasmids / drug effects Plasmids / genetics Promoter Regions, Genetic Protozoan Proteins* Recombinant Fusion Proteins / drug effects Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Repressor Proteins / chemistry Repressor Proteins / genetics Salmonella typhimurium / genetics Salmonella typhimurium / immunology Vaccines, Synthetic* |
| IF | 2.984 |
| 引用数 | 26 |
| WOS 分野 | GENETICS & HEREDITY |
| リソース情報 | |
| 原核生物(大腸菌) | pVUB3 |