Pm promoter expression mutants and their use in broad-host-range RK2 plasmid vectors
| Autor: | Janet Martha Blatny, Trygve Brautaset, Hanne C. Winther-Larsen, Bjørnar Valand, Svein Valla |
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| Rok vydání: | 2000 |
| Předmět: |
Mutant
Genetic Vectors Molecular Sequence Data Bioengineering Biology medicine.disease_cause Applied Microbiology and Biotechnology DNA-binding protein Plasmid Bacterial Proteins medicine Escherichia coli Point Mutation RK2 plasmid Replicon Promoter Regions Genetic Regulation of gene expression Genetics Base Sequence Point mutation Gene Expression Regulation Bacterial DNA-Binding Proteins Pseudomonas aeruginosa Trans-Activators Biotechnology Plasmids |
| Zdroj: | Metabolic engineering. 2(2) |
| ISSN: | 1096-7176 |
| Popis: | By coupling the Pm/xylS promoter system to minimal replicons of the broad-host-range plasmid RK2 we recently showed that such vectors are useful for both high- and low-level inducible expression of cloned genes in gram-negative bacteria. In this report, we extend this potential by identifying point mutations in or near the -10 transcriptional region of Pm. Point mutations leading to gene-independent enhancements of expression levels of the induced state or reduced background expression levels were identified using Escherichia coli as a host. By combining these mutations an additive effect in expression levels from the constructed Pm was observed. The highest induced expression level was obtained by inserting an E. coli consensus sigma70 - 10 recognition region. Most of the remaining activities in the reduced-background mutations appeared to originate from a transcriptional start site other than Pm. The effects of some of these mutations were also analyzed in Pseudomonas aeruginosa and were found to act similarly, but less pronounced in this host. |
| Databáze: | OpenAIRE |
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