Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
2090806 | Journal of Microbiological Methods | 2008 | 4 Pages |
In this study we developed an electrotransformation method for use with the Gram-positive bacterium Paenibacillus larvae—a deadly pathogen of honey bees. Combining multiple Bacillus electrotransformation methods to generate an initial protocol, we then optimized the following parameters for use with P. larvae: cell density of culture at harvest time, contents of the washing/electroporation solution, field strength of the electrical pulse, recovery growth medium, and recovery time period. With the optimized method, we achieved an average transformation efficiency of 1.9 × 105 transformants/μg DNA. The method is substantially different from the only other electrotransformation method for a Paenibacillus species found in the literature. This work should facilitate the study of the several previously discovered natural plasmids of P. larvae, and is a step toward developing a genetic system for this species.