| RRC ID |
33473
|
| 著者 |
Guzman LM, Belin D, Carson MJ, Beckwith J.
|
| タイトル |
Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter.
|
| ジャーナル |
J Bacteriol
|
| Abstract |
We have constructed a series of plasmid vectors (pBAD vectors) containing the PBAD promoter of the araBAD (arabinose) operon and the gene encoding the positive and negative regulator of this promoter, araC. Using the phoA gene and phoA fusions to monitor expression in these vectors, we show that the ratio of induction/repression can be 1,200-fold, compared with 50-fold for PTAC-based vectors. phoA expression can be modulated over a wide range of inducer (arabinose) concentrations and reduced to extremely low levels by the presence of glucose, which represses expression. Also, the kinetics of induction and repression are very rapid and significantly affected by the ara allele in the host strain. Thus, the use of this system which can be efficiently and rapidly turned on and off allows the study of important aspects of bacterial physiology in a very simple manner and without changes of temperature. We have exploited the tight regulation of the PBAD promoter to study the phenotypes of null mutations of essential genes and explored the use of pBAD vectors as an expression system.
|
| 巻・号 |
177(14)
|
| ページ |
4121-30
|
| 公開日 |
1995-7-1
|
| DOI |
10.1128/jb.177.14.4121-4130.1995
|
| PMID |
7608087
|
| PMC |
PMC177145
|
| MeSH |
Alkaline Phosphatase / biosynthesis
Alkaline Phosphatase / genetics
AraC Transcription Factor
Arabinose / metabolism*
Bacterial Proteins*
Base Sequence
Cloning, Molecular / methods*
Enzyme Induction
Enzyme Repression
Escherichia coli / genetics*
Escherichia coli Proteins
Gene Expression Regulation, Enzymologic*
Genes, Lethal / genetics
Genetic Vectors / genetics*
Molecular Sequence Data
Mutation
Operon / genetics
Phosphotransferases (Alcohol Group Acceptor) / genetics
Promoter Regions, Genetic*
Protein Engineering
Recombinant Fusion Proteins / biosynthesis
Repressor Proteins / genetics
Transcription Factors*
|
| IF |
3.006
|
| 引用数 |
3533
|
|
WOS 分野
|
MICROBIOLOGY
|
| リソース情報 |
| 原核生物(大腸菌) |
pBAD18
pBAD18-Cm
pBAD18-kan
pBAD18s
pBAD22A
pBAD24
pBAD28
pBAD30
pBAD33 |
