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bioRxiv · 10.1101/2020.03.03.974816

Efficient Dual-Negative Selection for Bacterial Genome Editing

Cianfanelli, F. R.·Cunrath, O.·Bumann, D.
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Abstract

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

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Cianfanelli, F. R., Cunrath, O., Bumann, D.. 2020-03-04. Efficient Dual-Negative Selection for Bacterial Genome Editing. https://doi.org/10.1101/2020.03.03.974816

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