Transformation of bacteria with plasmid DNA by electroporation

✍ Scribed by Stefan Fiedler; Reinhard Wirth

• Publisher: Elsevier Science
• Year: 1988
• Tongue: English
• Weight: 565 KB
• Volume: 170
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(88)90086-3

✦ Synopsis

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.