Rapid and Highly Efficient Method for Scarless Mutagenesis within the Salmonella enterica Chromosome

Autor(en): Blank, Kathrin
Hensel, Michael 
Gerlach, Roman G.
Stichwörter: ESCHERICHIA-COLI; GENE; IDENTIFICATION; Multidisciplinary Sciences; PATHOGENICITY; PHOP; PURIFICATION; RECOMBINATION; Science & Technology - Other Topics; SECRETION SYSTEM; TYPHIMURIUM; VIRULENCE
Erscheinungsdatum: 2011
Herausgeber: PUBLIC LIBRARY SCIENCE
Journal: PLOS ONE
Volumen: 6
Ausgabe: 1
Zusammenfassung: 
Direct manipulation of bacterial chromosomes by recombination-based techniques has become increasingly important for both cognitive and applied research. Here we demonstrate, for the first time, the combination of the Red recombinase system with I-SceI endonuclease-based selection of successful recombinants after electroporation with short synthetic olignucleotides. We show the generation of scarless gene knockouts as well as site-directed mutagenesis using the Salmonella virulence-associated two component signaling system PhoPQ. The presented approach is very versatile for generating in-frame deletions, point mutations or insertions within bacterial chromosomes.
ISSN: 19326203
DOI: 10.1371/journal.pone.0015763

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