Abstract
We have been succeeded in rice Waxy gene targeting. The genomic structure of our Waxy knock-out is expected to be substitution of theWaxy (Hmr) gene with a strong transcriptional stop element for a small segment within the Waxy1st intron. We have reported the preliminary characterization of 6 independent hetrozygousely targeted fertile lines (2002 Annual Meeting of JSPP). In order to examine whether precise homologous recombination events had occurred, we have performed detailed Southern blot analysis of the Waxy and its flanking regions of 35 kb and sequenced the entire 17 kb Waxyregion in the targeted T1 homozygous plants. Although it has been reported that undesirable ectopic recombination and ectopic integration of the transgene are often accompanied with gene targeting, no such events can be detected in our targeted lines. We can conclude that precise somatic homologous recombination events have occurred repeatedly at the Waxylocus.
