Development of Methods for the Target-Specific Protein Elimination in Plants

Resumo

Here we present the method for the target-specific elimination of certain intracellular proteins in plants using the ubiquitin-proteasome system. We modified the E3 ubiquitin ligase Chip of A. thaliana to obtain two variants carrying the deletions at the N-terminus and the GFP recognition domain. The interaction of the GFP protein and the chimeric ubiquitin ligase was confirmed via yeast two-hybrid assay. Fluorescence microscopy and fluorimetry showed that, when an infiltration of the gfp-expressing N. benthamiana plants was performed with agrobacteria carrying the hybrid E3 gene of the ubiquitin ligase Chip with the GFP recognition domain, a significant decrease in the fluorescence was observed for both variants: carrying the N-terminal deletion of 100 amino acids or the deletion of 140 amino acids.