AI Article Synopsis
- This study presents a new genetic mutation strategy using toxins that works well with clinical strains of E. coli and Salmonella without needing specific adjustments for each strain.
- The system offers better selection strength compared to previous methods, making it easier to conduct genetic experiments in various bacteria.
- It allows for allele transfers between E. coli strains without phage and can be adapted for use in other types of bacteria, addressing the limitations of current genetic tools.
Article Abstract
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.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4513841 | PMC |
| http://dx.doi.org/10.1093/nar/gkv248 | DOI Listing |
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