A protocol for transient gene expression in Arabidopsis thaliana protoplasts isolated from cell suspension cultures

Abstract

Isolation and maintenance of an Arabidopsis thaliana cell suspension culture, protoplast preparation from these cells and plasmid DNA transfection into the protoplasts by the polyethyleneglycol procedure are described. In the reported examples, the DNA contained chimaeric genes consisting on the coding region of the β-glucuronidase (GUS) reporter gene, fused to the cauliflower mosaic virus 35S promoter or to the promoter from the Arabidopsis elongation factor EF-1 α gene. Following DNA uptake into protoplasts, maximal GUS activity was observed after 50 to 60 h. The measured GUS activity indicated high transient expression of the tested chimaeric genes