Transformation of bacteria with plasmid DNA by electroporation.
The possibility of electric field-mediated transformation ("electroporation") of a gram-positive bacterium (Enterococcus faecalis) and two gram-negative bacteria (Escherichia coli and Pseudomonas putida) with plasmid DNA was investigated. E. faecalis protoplasts could be transformed by electroporation with a transformation frequency of 10(4) to 10(5) transformants/micrograms plasmid. Untreated--i.e., washed--cells of E. coli could be transformed with rates of 1 X 10(5) transformants/micrograms plasmid DNA. Transformation rates for P. putida cells were up to 3 X 10(4) if the method developed for E. coli was used. Detailed protocols for these systems, including the results of various optimization experiments, are given.
D004269:DNA, Bacterial; D004274:DNA, Recombinant; D004560:Electricity; D013293:Enterococcus faecalis / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D006090:Gram-Negative Bacteria / Q000235:genetics*; D006094:Gram-Positive Bacteria / Q000235:genetics*; D010957:Plasmids; D011523:Protoplasts; D011549:Pseudomonas / Q000235:genetics; D014169:Transformation, Bacterial
D004269:DNA, Bacterial; D004274:DNA, Recombinant