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- W1983996879 abstract "<ns4:p>Genetic manipulation in enterohemorrhagic<ns4:italic>E. coli</ns4:italic>O157:H7 is currently restricted to recombineering, a method that utilizes the recombination system of bacteriophage lambda, to introduce gene replacements and base changes<ns4:italic>inter alia</ns4:italic>into the genome. Bacteriophage 933W is a prophage in<ns4:italic>E. coli</ns4:italic>O157:H7 strain EDL933, which encodes the genes (<ns4:italic>stx2AB</ns4:italic>) for the production of Shiga toxin which is the basis for the potentially fatal Hemolytic Uremic Syndrome in infected humans. We replaced the<ns4:italic>stx2AB</ns4:italic>genes with a kanamycin cassette using recombineering. After induction of the prophage by ultra-violet light, we found that bacteriophage lysates were capable of transducing to wildtype, point mutations in the lactose, arabinose and maltose genes. The lysates could also transduce tetracycline resistant cassettes. Bacteriophage 933W is also efficient at transducing markers in<ns4:italic>E. coli</ns4:italic>K-12. Co-transduction experiments indicated that the maximal amount of transferred DNA was likely the size of the bacteriophage genome, 61 kB. All tested transductants, in both<ns4:italic>E. coli</ns4:italic>K-12 and O157:H7, were kanamycin-sensitive indicating that the transducing particles contained host DNA.</ns4:p>" @default.
- W1983996879 created "2016-06-24" @default.
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- W1983996879 date "2013-01-10" @default.
- W1983996879 modified "2023-10-03" @default.
- W1983996879 title "High efficiency generalized transduction in Escherichia coli O157:H7" @default.
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- W1983996879 doi "https://doi.org/10.12688/f1000research.2-7.v1" @default.
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