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 |
Zur Langanzeige
Seitenaufrufe
3
Letzte Woche
0
0
Letzter Monat
0
0
geprüft am 18.05.2024