<p style="text-align: justify;">Event 98140 was produced via <i>Agrobacterium</i>-mediated transformation of immature maize embryos using the plasmid vector PHP24279 (containing the <i>gat4621</i> and <i>zm-hra</i> gene cassettes). Immature embryos of maize were aseptically removed from the developing caryopsis 9 - 10 days after pollination and infected with <i>Agrobacterium tumefaciens</i> strain LBA4404 containing plasmid PHP24279, essentially as described in Zhao <i>et al</i>., (2001). After 6 days of embryo and <i>Agrobacterium</i> co-cultivation on solid culture medium with no selection, the embryos were transferred to selective medium that was stimulatory to maize somatic embryogenesis containing glyphosate for selection of cells expressing the <i>gat4621</i> transgene. The medium also contained carbenicillin to kill any remaining <i>Agrobacterium</i>.</p>

<p style="text-align: justify;">After two weeks on the selective medium, healthy, growing calli that demonstrated resistance to glyphosate were identified. The putative transgenic calli were continually transferred to fresh selection medium for further growth until the start of the regeneration process. The presence of both transgenes was confirmed using PCR analysis, and regenerated whole transgenic plants (T0) were transferred to the greenhouse where they were evaluated for glyphosate and ALS-inhibitor herbicide tolerance.</p>
