Electroporation and stable maintenance of plasmid DNAs in a biocontrol strain of Pseudomonas syringae

被引:10
作者
Bassett, CL [1 ]
Janisiewicz, WJ [1 ]
机构
[1] ARS, USDA, Appalachian Fruit Res Stn, Kearneysville, WV 25430 USA
关键词
broad host range plasmids; methylation deficiency; plasmid stability; transformation efficiency;
D O I
10.1023/A:1022394716305
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Transformation efficiencies as high as 10(7) transformants mug(-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.
引用
收藏
页码:199 / 203
页数:5
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