Transformation of Bacillus amyloliquefaciens by electroporation.
A method for transformation of whole Bacillus amyloliquefaciens cells by electroporation was developed. The procedure is as efficient as the protoplast transformation method, resulting in up to 10(5) transformants/micrograms plasmid DNA, but requires less effort and time. Cells for electroporation were grown to late exponential phase in a rich medium supplemented with 0.25 M sucrose, washed with and resuspended in 0.25 M sucrose, 1 mM HEPES, 1 mM MgCl2, 10% (v/v) glycerol, pH 7.0, at 3-5 x 10(10) cells/ml for storage at -80 degrees C. The highest transformation frequency was obtained at 7.5 kV/cm with a 25 microF capacitor. The transformation efficiency increased linearly with DNA concentration at least over the range 10 ng-12.5 micrograms/ml. Transformations with ligated DNA and of industrial strains were also successful. In addition, B. subtilis cells treated as above could be transformed by electroporation, resulting in 10(4) transformants/micrograms DNA at 12.5 kV/cm.
Vehmaanperä
J
J
Research Laboratories, Alko Ltd, Helsinki, Finland.
D001407:Bacillus / Q000235:genetics* / Q000254:growth & development; D004269:DNA, Bacterial / Q000302:isolation & purification; D004563:Electrochemistry; D010957:Plasmids; D014169:Transformation, Bacterial
D004269:DNA, Bacterial