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

<p style="text-align: justify;">Southern blot analysis of MON810 genomic DNA indicated the incorporation of a single copy of the truncated <i>cry1Ab</i> gene, together with the enhanced CaMV 35S (E35S) promoter and hsp70 leader sequences. The NOS 3&#39; termination signal, present in plasmid PV-ZMBK07, was not integrated into the host genome but was lost through a 3&#39; truncation of the gene cassette. The native Cry1Ab protein (HD-1) has a molecular weight of 131 kD while the inserted, plant expressed <i>cry1Ab</i> gene codes for a truncated protein with a molecular weight of 91 kD, as confirmed by Western blot analysis of MON810 tissue extracts. Evidence was provided that no plasmid backbone sequences from the plasmid PV-ZMGT10 were integrated into the MON810 genome. Further Southern blot analysis indicated that the genes for glyphosate tolerance (CP4 EPSPS) and antibiotic resistance (<i>neo</i>) were not transferred to line MON 810 and the absence of the CP4 EPSPS and <i>gox</i> gene products was also confirmed by Western blotting. The CP4 EPSPS and GOX protein encoding genes were presumed to have been inserted into the initial transformant at a separate genetic loci from the <i>cry1Ab</i> gene and then subsequently lost through segregation during the crossing events leading to line MON810.</p>

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

<p style="text-align: justify;">Segregation and stability data were consistent with a single site of insertion of the <i>cry1Ab</i> gene into the MON810 genome. The stability of the insertion was demonstrated through multiple generations of crossing. MON810 was derived from the third generation of backcrossing and stable integration of the single insert was demonstrated through all three generations by Southern Blot analysis.</p>

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

<p style="text-align: justify;">The synthetic <i>cry1Ab</i> gene was linked to a strong constitutive promoter and modified for maximum expression in corn. The amino acid sequence of the toxin expressed in the modified corn was found to be identical to that occurring naturally, and equivalent to that produced for use as the biopesticide that is widely used by the organic food industry. Average protein expression, as measured in samples obtained from field trials at six locations, was 9.35 &micro;g/g (fresh weight) in leaves and 0.31 &micro;g/g (f.w.) in seeds. The concentration of expressed toxin, as determined from a single sample obtained from one site, was 4.15 &micro;g/g (f.w.) in the whole plant and 0.09 &micro;g/g (f.w.) in pollen. Protein expression ranged from 7.93 to 10.34 &micro;g/g (f.w.) in leaves, from 0.19 to 0.39 &micro;g/g (f.w.) in grain, and from 3.65 to 4.65 &micro;g/g (f.w.) in the whole plant. Protein expression declined over the growing season as indicated by the Cry1Ab protein concentrations in leaves assayed over the growing season. 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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