Efficient dual-negative selection for bacterial genome editing.

ABSTRACT:

Background

Gene editing is key for elucidating gene function. Traditional methods, such as consecutive single-crossovers, have been widely used to modify bacterial genomes. However, cumbersome cloning and limited efficiency of negative selection often make this method slower than other methods such as recombineering.

Results

Here, we established a time-effective variant of consecutive single-crossovers. This 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. Adapted versions enabled efficient gene editing also in Pseu

SUBMITTER: Cianfanelli FR 

PROVIDER: S-EPMC7245781 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature