A robust one-step recombineering system for enterohemorrhagic escherichia coli

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dc.contributor.authorPeng, Lang
dc.contributor.authorDumevi, Rexford Mawunyo
dc.contributor.authorChitto, Marco
dc.contributor.authorHaarmann, Nadja
dc.contributor.authorBerger, Petya
dc.contributor.authorKoudelka, Gerald
dc.contributor.authorSchmidt, Herbert
dc.contributor.authorMellmann, Alexander
dc.contributor.authorDobrindt, Ulrich
dc.contributor.authorBerger, Michael
dc.date.accessioned2024-09-03T14:03:48Z
dc.date.available2024-09-03T14:03:48Z
dc.date.issued2022de
dc.description.abstractEnterohemorrhagic Escherichia coli (EHEC) can cause severe diarrheic in humans. To improve therapy options, a better understanding of EHEC pathogenicity is essential. The genetic manipulation of EHEC with classical one-step methods, such as the transient overexpression of the phage lambda (λ) Red functions, is not very efficient. Here, we provide a robust and reliable method for increasing recombineering efficiency in EHEC based on the transient coexpression of recX together with gam, beta, and exo. We demonstrate that the genetic manipulation is 3–4 times more efficient in EHEC O157:H7 EDL933 Δstx1/2 with our method when compared to the overexpression of the λ Red functions alone. Both recombineering systems demonstrated similar efficiencies in Escherichia coli K-12 MG1655. Coexpression of recX did not enhance the Gam-mediated inhibition of sparfloxacin-mediated SOS response. Therefore, the additional inhibition of the RecFOR pathway rather than a stronger inhibition of the RecBCD pathway of SOS response induction might have resulted in the increased recombineering efficiency by indirectly blocking phage induction. Even though additional experiments are required to unravel the precise mechanistic details of the improved recombineering efficiency, we recommend the use of our method for the robust genetic manipulation of EHEC and other prophage-carrying E. coli isolates.en
dc.identifier.swb1818275244
dc.identifier.urihttps://hohpublica.uni-hohenheim.de/handle/123456789/16591
dc.identifier.urihttps://doi.org/10.3390/microorganisms10091689
dc.language.isoengde
dc.rights.licensecc_byde
dc.source2076-2607de
dc.sourceMicroorganisms; Vol. 10, No. 9 (2022) 1689de
dc.subjectEnterohemorrhagic escherichia coli
dc.subjectRecombineering
dc.subjectSOS response
dc.subject.ddc570
dc.titleA robust one-step recombineering system for enterohemorrhagic escherichia colien
dc.type.diniArticle
dcterms.bibliographicCitationMicroorganisms, 10 (2022), 9, 1689. https://doi.org/10.3390/microorganisms10091689. ISSN: 2076-2607
dcterms.bibliographicCitation.issn2076-2607
dcterms.bibliographicCitation.issue9
dcterms.bibliographicCitation.journaltitleMicroorganisms
dcterms.bibliographicCitation.volume10
local.export.bibtex@article{Peng2022, url = {https://hohpublica.uni-hohenheim.de/handle/123456789/16591}, doi = {10.3390/microorganisms10091689}, author = {Peng, Lang and Dumevi, Rexford Mawunyo and Chitto, Marco et al.}, title = {A Robust One-Step Recombineering System for Enterohemorrhagic Escherichia coli}, journal = {Microorganisms}, year = {2022}, volume = {10}, number = {9}, }
local.export.bibtexAuthorPeng, Lang and Dumevi, Rexford Mawunyo and Chitto, Marco et al.
local.export.bibtexKeyPeng2022
local.export.bibtexType@article

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