Development of a reinforced Ti-eviction plasmid useful for construction of Ti plasmid-free Agrobacterium strains

For curing stable Ti plasmids from Agrobacterium strains, we developed a new Ti-eviction plasmid. The plasmid contains two antitoxin genes, which originated from two toxin-antitoxin systems in Ti plasmids. The plasmid facilitated production of Ti-less cells in seven tested strains, and was removed easily by simple counter-selection from the cells.

► We constructed a new Ti-eviction plasmid containing antitoxin genes of TA systems.
► Introduction of the new plasmid induced Ti-free cells at high frequency.
► The new plasmid was removed easily by simple counter-selection from the cells.