Efficient Dual-Negative Selection for Bacterial Genome Editing

• DOI: 10.1101/2020.03.03.974816
• Publication Date: 2020-03-05T06:45:15Z
• Authors: Francesca Romana ...; Olivier Cunrath; Dirk Bumann
• Keywords: Salmonella enterica; Strain (injury)

Abstract We describe a versatile method for chromosomal gene editing based on classical consecutive single-crossovers. The exploits rapid plasmid construction using Gibson assembly, convenient E. coli donor strain, and efficient dual-negative selection improved suicide vector resolution. used this to generate in frame deletions, insertions point mutations Salmonella enterica with limited hands-on time. Similar strategies allowed also Pseudomonas aeruginosa multi-drug-resistant (MDR) Escherichia clinical isolates.