<p style="text-align: justify;"><b>The Introduced DNA</b></p>

<p style="text-align: justify;">Southern blot analysis of MON809 genomic DNA indicated one integrated DNA segment, of approximately 23Kb DNA, which included a complete and a partial copy of the <i>cry1Ab</i> gene, two complete copies of the CP4 EPSPS encoding gene, a partial <i>gox</i> gene and the NPT II encoding antibiotic resistance marker gene. Additional Southern blot analyses were performed to demonstrate that additional plasmid backbone DNA sequences from PV-ZMBK07 were not integrated into the host genome.</p>

<p style="text-align: justify;"><b>Genetic Stability of the Introduced Trait</b></p>

<p style="text-align: justify;">Segregation analysis over five generations of crossing indicated that the <i>cry1Ab</i> gene was stably integrated and segregated according to Mendelian rules of inheritance.</p>

<p style="text-align: justify;"><b>Expressed Material</b></p>

<p style="text-align: justify;">Western blot analysis of protein extracts from MON809 demonstrated the presence of an immunoreactive protein of the predicted size. Lower molecular weight species corresponding to the predicted size of the truncated cry1Ab gene were not detected, indicating that this partial gene sequence is not expressed. The expression of Cry1Ab protein was evaluated using enzyme linked immunosorbent assay (ELISA) of tissue samples obtained from plants grown at 6 locations. The average levels of expressed Cry1Ab protein were: 1.63 &plusmn; 0.75 &micro;g/g fresh weight (fwt) in leaves, 0.55 &plusmn; 0.23 &micro;g/g fwt in grain and 1.23 &plusmn; 0.5 &micro;g/g fwt in the whole plant. Cry1Ab protein was not detected in pollen and the levels expressed in leaves declined over the growing season. Similarly, CP4 EPSPS gene activity, used as a selectable marker conferring tolerance to glyphosate, was quantified by testing for EPSPS protein levels and averaged results were as follows: 21.68 &micro;g/g fwt in leaves, 9.41 &micro;g/g fwt in grain and 1.6 &micro;g/g fwt in the whole plant. Data were provided showing that the level of expression of CP4 EPSPS was too low to confer significant glyphosate resistance in the field. Expression of the second gyphosate tolerance gene, <i>gox</i>, which encodes the enzyme glyphosate oxidase, could not be detected by Western blot analysis. The Cry1Ab protein was shown to degrade readily in the environment. The plant expressed protein had DT50 and DT90 values (time to degrade to 50% and 90 % of the original bioactivity) of 2 and 15 days respectively.</p>

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