Patent ID: 7126041

Claim:
A method for the targeted insertion of a nucleotide of interest into a specific chromosomal site within a plant cell, said method comprising the steps of: (a) providing a plant cell having a heterologous target site on a chromosome thereof, wherein said target site is flanked only on one side by a single recombination site, which single recombination site is recognized by a site-specific recombinase enzyme; and then (b) transforming said plant cell with an Agrobacterium transformation vector carrying a nucleotide sequence of interest, wherein said nucleotide sequence of interest is flanked by a pair of identical recombination sites, one on each side thereof, that correspond to the single recombination site of said target site, so that said nucleotide of interest (i) is randomly inserted into a chromosome of said plant cell, (ii) generates an excision circle therefrom, and then (iii) is inserted into said chromosome at said target site; wherein said transforming step is carried out in the presence of a site-specific recombinase effective to carry out recombination at said recombination site and insert said nucleotide of interest into said chromosome at said target site.