Transformation of the Gram-positive honey bee pathogen, Paenibacillus larvae, by electroporation.
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 1.9x10(5) transformants/mug 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.
Murray
K Daniel
KD
USDA-ARS, Honey Bee Research Unit, Kika de la Garza Subtropical Agricultural Center, 2413 E. Hwy 83, Weslaco, TX 78596, United States. keith.murray@ars.usda.gov
Aronstein
Katherine A
KA
D000818:Animals; D001516:Bees / Q000382:microbiology*; D003470:Culture Media; D018274:Electroporation / Q000379:methods*; D016937:Gram-Positive Endospore-Forming Rods / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D007814:Larva / Q000382:microbiology; D010957:Plasmids / Q000235:genetics; D013171:Spores, Bacterial; D014169:Transformation, Bacterial
D003470:Culture Media