High efficiency generalized transduction in Escherichia coli O157:H7 [v1; ref status: indexed, http://f1000r.es/8f]

Genetic manipulation in enterohemorrhagic O157:H7 is currently E. coli restricted to recombineering, a method that utilizes the recombination system of bacteriophage lambda, to introduce gene replacements and base changes into the genome. Bacteriophage 933W is a prophage in inter alia E. coli O157:H7 strain EDL933, which encodes the genes ( ) for the production stx2AB of Shiga toxin which is the basis for the potentially fatal Hemolytic Uremic Syndrome in infected humans. We replaced the genes with a stx2AB 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 K-12. E. coli 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 K-12 and O157:H7, were kanamycin-sensitive E. coli indicating that the transducing particles contained host DNA. Martin G Marinus ( ) Corresponding author: [email protected] Marinus MG, Poteete AR (2013) High efficiency generalized transduction in O157:H7 [v1; ref status: How to cite this article: Escherichia coli indexed, ] 2013, :7 (doi: 10.12688/f1000research.2-7.v1) http://f1000r.es/8f F1000Research 2 This is an open access article distributed under the terms of the , which permits unrestricted Copyright: Creative Commons Attribution Licence use, distribution, and reproduction in any medium, provided the original work is properly cited. Data associated with the article are available under the terms of the (CC0 1.0 Public domain dedication). Creative Commons Zero "No rights reserved" data waiver This work was supported by NIH grant GM063790 to MGM. Grant information: The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. No competing interests were disclosed. Competing Interests: 10 Jan 2013, :7 (doi: 10.12688/f1000research.2-7.v1) First Published: 2 21 Jan 2013, :7 (doi: 10.12688/f1000research.2-7.v1) First Indexed: 2 Referees v1 published 10 Jan 2013 1 2 3 report 1 report report 10 Jan 2013, :7 (doi: 10.12688/f1000research.2-7.v1) First Published: 2 10 Jan 2013, :7 (doi: 10.12688/f1000research.2-7.v1) Latest Published: 2 v1 Page 1 of 7 F1000Research 2013, 2:7 Last updated: 07 AUG 2013