PT  - JOURNAL ARTICLE
AU  - Zuo, Fanglei
AU  - Zeng, Zhu
AU  - Hammarström, Lennart
AU  - Marcotte, Harold
TI  - Inducible Plasmid Self-Destruction (IPSD) assisted genome engineering in lactobacilli and bifidobacteria
AID  - 10.1101/649921
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 649921
4099  - http://biorxiv.org/content/early/2019/05/24/649921.short
4100  - http://biorxiv.org/content/early/2019/05/24/649921.full
AB  - Genome engineering is essential for application of synthetic biology in probiotics including lactobacilli and bifidobacteria. Several homologous recombination system-based mutagenesis tools have been developed for these bacteria but still, have many limitations in different species or strains. Here we developed a genome engineering method based on an inducible self-destruction plasmid delivering homologous DNA into bacteria. Excision of the replicon by induced recombinase facilitates selection of homologous recombination events. This new genome editing tool called Inducible Plasmid Self-Destruction (IPSD) was successfully used to perform gene knock-out and knock-in in lactobacilli and bifidobacteria. Due to its simplicity and universality, the IPSD strategy may provide a general approach for genetic engineering of various bacterial species.