ArticleApplied and environmental microbiology2011
Isolation of generalized transducing bacteriophages for uropathogenic strains of Escherichia coli.
Article in Applied and environmental microbiology, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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29 citing papers in PubMed, 41 citations in OpenAlex.
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- Bacterial and Bacteriophage Antibiotic Resistance in Marine Bathing Waters in Relation to Rivers and Urban Streams.Frontiers in microbiology · 2021Article
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- Genetic cargo and bacterial species set the rate of vesicle-mediated horizontal gene transfer.Scientific reports · 2017Article
- Bacteriophages Contribute to the Spread of Antibiotic Resistance Genes among Foodborne Pathogens of theFrontiers in microbiology · 2017Review
- Transposon-Sequencing Analysis Unveils Novel Genes Involved in the Generation of Persister Cells in Uropathogenic Escherichia coli.Antimicrobial agents and chemotherapy · 2016Article
- A set of powerful negative selection systems for unmodified Enterobacteriaceae.Nucleic acids research · 2015Article
- Recombinant transfer in the basic genome of Escherichia coli.Proceedings of the National Academy of Sciences of the United States of America · 2015Article
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6 authors at 1 institution in 1 country.
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Abstract
The traditional genetic procedure for random or site-specific mutagenesis in Escherichia coli K-12 involves mutagenesis, isolation of mutants, and transduction of the mutation into a clean genetic background. The transduction step reduces the likelihood of complications due to secondary mutations. Though well established, this protocol is not tenable for many pathogenic E. coli strains, such as uropathogenic strain CFT073, because it is resistant to known K-12 transducing bacteriophages, such as P1. CFT073 mutants generated via a technique such as lambda Red mutagenesis may contain unknown secondary mutations. Here we describe the isolation and characterization of transducing bacteriophages for CFT073. Seventy-seven phage isolates were acquired from effluent water samples collected from a wastewater treatment plant in Madison, WI. The phages were differentiated by a host sensitivity-typing scheme with a panel of E. coli strains from the ECOR collection and clinical uropathogenic isolates. We found 49 unique phage isolates. These were then examined for their ability to transduce antibiotic resistance gene insertions at multiple loci between different mutant strains of CFT073. We identified 4 different phages capable of CFT073 generalized transduction. These phages also plaque on the model uropathogenic E. coli strains 536, UTI89, and NU14. The highest-efficiency transducing phage, ΦEB49, was further characterized by DNA sequence analysis, revealing a double-stranded genome 47,180 bp in length and showing similarity to other sequenced phages. When combined with a technique like lambda Red mutagenesis, the newly characterized transducing phages provide a significant development in the genetic tools available for the study of uropathogenic E. coli.
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