FACTORS INFLUENCING TRANSIENT GENE-EXPRESSION IN BEAN (PHASEOLUS-VULGARIS L) USING AN ELECTRICAL PARTICLE-ACCELERATION DEVICE

The parameters influencing transient expression of the beta-glucuronidase gene in bean embryonic axes, cotyledons, apical meristems and leaves were evaluated after gene delivery with an electrical particle acceleration device. A calcium-spermidine procedure for coating gold particles with DNA resulted in higher levels of GUS expression with lower concentrations of gold particles compared with a calcium phosphate procedure. The DNA concentration, distance between the discharge chamber and the retaining screen and the vacuum in the apparatus also influenced gene delivery. Sections prepared for light and electron microscopy showed the localisation, within target cells, of gold particles used to deliver the DNA. Immunolocalization of foreign gene expression within cells confirmed an even distribution of gene product throughout the cell cytoplasm.