Electroporation and stable maintenance of plasmid DNAs in a biocontrol strain of Pseudomonas syringae.
Transformation efficiencies as high as 10(7) transformants microg(-1) DNA have been previously reported for pseudomonads using electroporation protocols established for E. coli with plasmid DNAs prepared from methylation proficient E. coli hosts. We report here a protocol for electroporation of plasmid DNAs into a biocontrol strain of Pseudomonas syringae which could not be electroporated by standard E. coli methods. Transformation efficiencies of 10(7) or higher were obtained with DNA recovered from initial P. syringae transformation or with DNA prepared from methylation deficient E. coli. Both plasmids used in this study were stably maintained in the absence of selection for at least 50 generations.
Bassett
Carole L
CL
USDA, Agricultural Research Service, Appalachian Fruit Research Station, 45 Wiltshire Road, Kearneysville, WV 25430, USA. cbassett@afrs.ars.usda.gov
Janisiewicz
Wojciech J
WJ
D000667:Ampicillin / Q000494:pharmacology; D018274:Electroporation / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods; D007612:Kanamycin / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics* / Q000493:pharmacokinetics*; D011549:Pseudomonas / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D011786:Quality Control; D014170:Transformation, Genetic
D007612:Kanamycin; D000667:Ampicillin