Genetic Transformation of the Model Quorum Sensing Bacterium by Electroporation.

BACKGROUND

The marine bacterium has been a model system for the study of bacterial quorum sensing and is increasingly recognized as a formidable aquatic animal pathogen. While genetically tractable, the study of this species in basic and applied research still relies upon laborious and time-consuming conjugation methods for plasmid DNA transformation.

METHODS

In this study, we developed an electroporation protocol using the most studied strain of this species, ATCC BAA-1116. An electroporation efficiency of up to 3 × 10 CFU/μg DNA was demonstrated using derived parameters (10 kV/cm, 400 Ω, 25 μF), which took cell growth phase at harvest, plasmid DNA amount, and recovery conditions into account. The electroporation protocol was tested using several different plasmids and with additional strains of and sister species .

RESULTS

Interestingly, of the eight other strains tested, only three others, which also happened to be the three most recent environmental isolates with the fewest number of laboratory passages, were amenable to electroporation-mediated transformation.

CONCLUSIONS

This electroporation protocol expands the tool set for studying and provides interesting insights into DNA transformation and uptake in this and related bacterial species.