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Abstract
Electric field-mediated gene transfer in mammalian cells (electrotransformation) depends
on the pulsing conditions (field intensity, pulse duration, number of pulses). The
effect of these parameters was systematically investigated using the transient expression
of the chloramphenicol acetyltransferase and the beta-galactosidase activities in
Chinese hamster ovary cells. Pulsing conditions inducing reversible permeabilization
of the cell plasma membrane are not sufficient to induce gene transfer. The plasmid
must be present during the electric pulse if it is to be transferred across the membrane
into the cytoplasm. Only the localized part of the cell membrane brought to the permeabilized
state by the external field is competent. Pulse duration plays a key role in the magnitude
of the transfer. The field induces a complex reaction between the membrane and the
plasmid that is accumulated at the cell interface by electrophoretic forces. This
leads to an insertion of the plasmid, which can then cross the membrane.