Development of an efficient electroporation method for rhizobacterial Bacillus mycoides strains

- An efficient electroporation method has been developed for rhizobacterial B. mycoides.
- The electroporation efficiency of B. mycoides EC18 was improved to 1.3 × 105 cfu/μg DNA.
- The electroporation protocol can be applied to other environmental B. mycoides strains.
- Restriction/modification is the main barrier for electroporation of this bacterium.

In order to develop a method for electroporation of environmental Bacillus mycoides strains, we optimized several conditions that affect the electroporation efficiency of this bacterium. By combining the optimized conditions, the electroporation efficiency of strain EC18 was improved to (1.3 ± 0.6) × 105 cfu/μg DNA, which is about 103-fold increase in comparison with a previously reported value. The method was further validated on various B. mycoides strains, yielding reasonable transformation efficiencies. Furthermore, we confirmed that restriction/modification is the main barrier for electroporation of this bacterium. To the best of our knowledge, this is the first systematic investigation of various parameters of electroporation of B. mycoides. The electroporation method reported will allow for efficient genetic manipulation of this bacterium.