Method for Bxb1-Mediated Site-Specific Integration In Planta

Gene targeting in plants through homologous recombination has been sparsely reported, although notable breakthroughs have been achieved in recent years (
1
). In particular, the use of zinc finger nucleases to promote homologous end joining has revived the promise that homologous gene targeting could someday become practical for plant genetic engineering (
2, 3
). An alternative and complementary approach that has progressed steadily over the years has been recombinase-mediated site-specific integration (
4
). In this approach, a first recombination site is introduced into the genome to serve as a target for inserting subsequent
DNA
. Here, we describe the method for generating the chromosomal target and the subsequent insertion of new DNA into the chromosomal target by Bxb1-mediated site-specific integration. This method would permit the comparison of different molecular constructs at the same genomic locations.