Electroporation Method for Gene Transfer
This method is based on the use of short electrical pulses of high field strength. These pulses increase the permeability of protoplast membrane and facilitates entry of DNA molecules into the cells, provided the DNA is in direct contact with the membrane. In view of this, for delivery of DNA to protoplasts, electroporation is one of the several routine techniques for efficient transformation. However, since regeneration from protoplasts is not always possible, cultured cells or tissue explants are often used. Consequently, it is important to test whether electroporation could transfer genes into walled cells. In most of these cases no proof of transformation was available.

Protoplasts in an ionic solution containing the vector DNA, are Fusion of plasmid-filled liposomes with isolated protoplasts as a method of plasmid uptake.suspended between the electrodes, electroporated and then plated as usual. Transformed colonies are selected as described earlier. Using method.Successful transfer of genes was achieved with the protoplasts of tobacco, petunia, maize, rice, wheat and sorghum. In most of these cases catgene associated with a suitable promoter sequence was transferred. Transformation frequencies can be further improved by (i) using field strength of 1.25 kV/cm, (ii) adding PEG after adding DNA, (iii) heat shocking protoplasts at 45°C for 5 minutes before adding DNA and (iv) by using linear instead of circular DNA.