Transient and Stable Expression of Foreign DNA Introduced into Plant Protoplasts by Electroporation

Electroporation utilizes high-voltage electric fields for cell permeabilization, This technique has been used for promoting the cellular uptake of exogenous molecules and macromolecules, including nucleotides, dyes, RNA, DNA, and even small proteins (1 -7 ). Electroporation′s useful attributes are its simplicity and its general effectiveness with a wide range of cell types. Because of its general efficacy, electroporation is becoming a valuable technique for the introduction of DNA into cell types that are resistant to transformation by other procedures (6 -8 ). In this chapter, we will outline procedures for introducing foreign DNAs into plant protoplasts by electroporation. DNA uptake is verified by the transient expression of chloramphenicol acetyltransferase (CAT), and by the recovery of plants stably transformed to kanamycin resistance through the introduction of the neomycin phosphotransferase gene.