Efficient electrotransformation of Enterococcus hirae with a new Enterococcus-Escherichia coli shuttle vector.
In the present study, an Enterococcus-Escherichia coli shuttle vector, pC3, was constructed that allows efficient transformation by electroporation of Enterococcus hirae ATCC9790. 5 x 10(6) transformants per microgram of plasmid DNA were obtained, using a commercial capacitor discharge device with an improved circuitry and a home-made electrode assembly, delivering pulses of 24 kV/cm across the cell suspension. The transformants were stable without selective pressure and plasmid DNA reisolated from transformed cells displayed no alterations in restriction enzyme analysis. Chromosomal DNA from E coli or E hirae, carried by pC3, was stably maintained in E hirae, making cloning and genetic manipulation in this organism feasible.
Solioz
M
M
Dept of Clinical Pharmacology, University of Berne, Switzerland.
Waser
M
M
D004269:DNA, Bacterial / Q000235:genetics*; D004566:Electrodes; D004926:Escherichia coli / Q000235:genetics*; D010957:Plasmids; D013291:Streptococcus / Q000235:genetics*; D014169:Transformation, Bacterial
D004269:DNA, Bacterial