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    High efficiency generalized transduction in Escherichia coli O157:H7

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    Authors
    Marinus, Martin G.
    Poteete, Anthony R.
    UMass Chan Affiliations
    Department of Microbiology and Physiological Systems
    Department of Biochemistry and Molecular Pharmacology
    Document Type
    Journal Article
    Publication Date
    2013-01-10
    Keywords
    Transduction, Genetic
    Genetics and Genomics
    Microbiology
    
    Metadata
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    Abstract
    Genetic manipulation in enterohemorrhagic E. coli O157:H7 is currently restricted to recombineering, a method that utilizes the recombination system of bacteriophage lambda, to introduce gene replacements and base changes inter alia into the genome. Bacteriophage 933W is a prophage in E. coli O157:H7 strain EDL933, which encodes the genes ( stx2AB) for the production of Shiga toxin which is the basis for the potentially fatal Hemolytic Uremic Syndrome in infected humans. We replaced the stx2AB 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 E. coli 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 E. coli K-12 and O157:H7, were kanamycin-sensitive indicating that the transducing particles contained host DNA.
    Source
    Marinus MG, Poteete AR. High efficiency generalized transduction in Escherichia coli O157:H7. F1000Res. 2013 Jan 10;2. doi:10.12688/f1000research.2-7.v1. Link to article on publisher's website
    DOI
    10.12688/f1000research.2-7.v1
    Permanent Link to this Item
    http://hdl.handle.net/20.500.14038/30052
    PubMed ID
    24327879
    Related Resources
    Link to article in PubMed
    Rights
    Copyright 2013 Marinus MG et al. This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted 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 Creative Commons Zero "No rights reserved" data waiver (CC0 1.0 Public domain dedication).
    ae974a485f413a2113503eed53cd6c53
    10.12688/f1000research.2-7.v1
    Scopus Count
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    UMass Chan Faculty and Researcher Publications

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