M. Nicolaisen et Gb. Poulsen, OPTIMIZATION OF POLYETHYLENE-GLYCOL MEDIATED TRANSIENT GENE-EXPRESSION IN PEA PROTOPLASTS, Plant cell, tissue and organ culture, 35(1), 1993, pp. 93-97
We have investigated factors influencing polyethylene glycol mediated
DNA uptake and beta-glucuronidase expression in pea (Pisum sativum L.)
protoplasts. It was found that for optimal beta-glucuronidase express
ion the molecular weight and concentration of polyethylene glycol shou
ld be 4000 and 20%, respectively. The amount of plasmid DNA should be
25 mug per 5 x 10(5) protoplasts in each treatment, and the concentrat
ion of Mg2+ in the transformation buffer should be 15 mM. The optimize
d protocol was applicable to all four pea cultivars tested.