Methods to find optimal integration sites within a plant genome are
provided. More particularly, a plant is transformed with a target site
having an expression cassette comprising a nucleotide sequence operably
linked to a promoter active in the plant. The target site is flanked by
non-identical recombination sites. Transformed protoplast, tissues, or
whole plants can be tested to determine the levels of activity of the
inserted gene. By comparison of cellular activities of the gene in
different insertion sites, preferred integration sites may be found
wherein the gene is expressed at high or acceptable levels. These plants
can then be utilized with subsequent retargeting techniques to replace
the nucleotide sequence with other genes or nucleotide sequences of
interest contained in a transfer cassette.