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.
References
Oct 5, 1975·Journal of Molecular Biology·N KlecknerD Botstein
Mar 23, 1968·Nature·M Meselson, R Yuan
Apr 29, 1998·Journal of Bacteriology·K C Murphy
Mar 12, 1999·Journal of Bacteriology·G PlunkettF R Blattner
Feb 24, 2001·Nature·N T PernaF R Blattner
Mar 10, 2001·The Journal of Clinical Investigation·M S Donnenberg, T S Whittam
Nov 14, 2002·Annual Review of Genetics·Donald L CourtLynn C Thomason
Jul 15, 1962·Proceedings of the National Academy of Sciences of the United States of America·E C HEATH, A D ELBEIN
Dec 16, 2003·BMC Molecular Biology·Kenan C Murphy, Kenneth G Campellone
Mar 26, 2004·Nature Reviews. Microbiology·James B KaperHarry L Mobley
Nov 2, 2004·Journal of Bacteriology·Jessica S TylerDavid I Friedman
Feb 2, 2006·FEMS Microbiology Letters·Kevin J SpearsDavid L Gally
Dec 13, 2006·Infection and Immunity·Theresa Deland Ho, Matthew K Waldor
Sep 11, 2007·DNA Repair·Anetta Nowosielska, M G Marinus
Citations
Apr 6, 2018·Journal of Medical Microbiology·Thujitha Thuraisamy, Patricia B Lodato
Apr 26, 2019·Microbial Drug Resistance : MDR : Mechanisms, Epidemiology, and Disease·Lekshmi NarendrakumarSabu Thomas
Apr 6, 2017·Frontiers in Microbiology·Lutz GeueStefanie A Barth
Jul 6, 2017·Frontiers in Microbiology·Anna ColavecchioLawrence D Goodridge
Jun 20, 2020·MicrobiologyOpen·Cinzia FinoAlexander Harms
Mar 7, 2021·Pharmaceuticals·Ahmad Y HassanHany Anany