Abstract
Oligonucleotide-directed gene targeting, which cause site-specific base change in the target gene through introducing the synthesized oligonuceotides directly into cells, was demonstrated. In plants, this method was only demonstrated in a chemically selectable gene with low efficiency. In this study, we established the assay system in order to search the effective condition of oligonucleotide-directed gene targeting in short period. We produced transgenic rice plants overexpressing mutated GUS gene (35S:mGUS). 35S:mGUS-introduced calli cells had GUS activity only in the case that they had predicted base change at an artificial stop codon. This mGUS repairing assay system took 8-10 days to compare the efficiency in various conditions. We are comparing an effective construct and amount of the oligonucleotides, as well as co-bombardment of plasmid DNA containing genes for DNA repairing enzymes. Improvement of the condition of oligonucleotide-directed gene targeting is now in progress.
