Electroporation and stable maintenance of plasmid DNAs in a biocontrol strain of Pseudomonas syringae |
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Authors: | Bassett Carole L Janisiewicz Wojciech J |
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Institution: | (1) USDA, Agricultural Research Service, Appalachian Fruit Research Station, 45 Wiltshire Road, Kearneysville, WV 25430, USA |
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Abstract: | Transformation efficiencies as high as 107 transformants g–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 107 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. |
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Keywords: | broad host range plasmids methylation deficiency plasmid stability transformation efficiency |
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