Electrotransformation of Streptococcus pyogenes with plasmid and linear DNA.
Electrotransformation was used to introduce both plasmid and linear DNA into Streptococcus pyogenes. The method was optimized using strain NZ131, for which transformation frequencies up to 10(7) per micrograms of plasmid DNA were obtained. A linear fragment of DNA, containing the streptokinase gene (ska) in which an internal fragment had been replaced with an erythromycin resistance gene (erm), was transformed into strain NZ131 with a frequency of 10(3) per micrograms DNA. The introduction of linear DNA into S. pyogenes by electrotransformation should be useful for future genetic analyses as well as targeted gene replacement.
Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City 73190.
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Simon
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Ferretti
D015139:Blotting, Southern; D004269:DNA, Bacterial / Q000235:genetics*; D004352:Drug Resistance, Microbial / Q000235:genetics; D004558:Electric Stimulation; D004917:Erythromycin / Q000494:pharmacology; D007700:Kinetics; D010957:Plasmids / Q000235:genetics*; D013297:Streptococcus pyogenes / Q000187:drug effects / Q000235:genetics*; D013300:Streptokinase / Q000235:genetics / Q000378:metabolism; D014169:Transformation, Bacterial / Q000235:genetics*
D004269:DNA, Bacterial; D004917:Erythromycin; D013300:Streptokinase