Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter
| Autor: | Luz-Maria Guzman, M. J. Carson, Jon Beckwith, Dominique Belin |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 1995 |
| Předmět: |
Arabinose
Escherichia coli/ genetics Operon Recombinant Fusion Proteins Recombinant Fusion Proteins/biosynthesis Genetic Vectors AraC Transcription Factor Molecular Sequence Data Repressor Biology ddc:616.07 Protein Engineering Microbiology Gene Expression Regulation Enzymologic Alkaline Phosphatase/biosynthesis/genetics chemistry.chemical_compound Plasmid Bacterial Proteins Genetic Vectors/ genetics Escherichia coli Arabinose/ metabolism Inducer Cloning Molecular Phage shock Promoter Regions Genetic Molecular Biology Regulation of gene expression Genes Lethal/genetics Repressor Proteins/genetics Base Sequence Escherichia coli Proteins PBAD promoter Alkaline Phosphatase Molecular biology Cloning Molecular/ methods Phosphotransferases (Alcohol Group Acceptor)/genetics Repressor Proteins Phosphotransferases (Alcohol Group Acceptor) chemistry Enzyme Induction Mutation Genes Lethal Enzyme Repression Operon/genetics Research Article Transcription Factors |
| Zdroj: | Journal of Bacteriology, Vol. 177, No 14 (1995) pp. 4121-4130 |
| ISSN: | 0021-9193 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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