A set of powerful negative selection systems for unmodified Enterobacteriaceae
| Autor: | Swaine L. Chen, Siyi Chen, Chiew Ling Lim, Varnica Khetrapal, Shazmina Rafee, Kurosh S. Mehershahi |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Salmonella
Bacterial Toxins medicine.disease_cause Negative selection Plasmid Enterobacteriaceae Transduction Genetic Genetics medicine Escherichia coli Humans Cloning Molecular Gene Cell Engineering Alleles Bacterial disease biology business.industry Salmonella enterica Modular design Chromosomes Bacterial biology.organism_classification Genetic Loci Mutation Methods Online business |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| Popis: | Creation of defined genetic mutations is a powerful method for dissecting mechanisms of bacterial disease; however, many genetic tools are only developed for laboratory strains. We have designed a modular and general negative selection strategy based on inducible toxins that provides high selection stringency in clinical Escherichia coli and Salmonella isolates. No strain- or species-specific optimization is needed, yet this system achieves better selection stringency than all previously reported negative selection systems usable in unmodified E. coli strains. The high stringency enables use of negative instead of positive selection in phage-mediated generalized transduction and also allows transfer of alleles between arbitrary strains of E. coli without requiring phage. The modular design should also allow further extension to other bacteria. This negative selection system thus overcomes disadvantages of existing systems, enabling definitive genetic experiments in both lab and clinical isolates of E. coli and other Enterobacteriaceae. |
| Databáze: | OpenAIRE |
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