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

<p style="text-align: justify;">Molecular analyses showed that maize lines T14 and T25 contained 3 and 1 copies, respectively, of the <i>pat</i> gene. Further analyses demonstrated that neither T14 nor T25 possessed a complete sequence of the antibiotic resistance encoding <i>bla</i> gene. Northern blot analyses showed that maize lines derived from T14 and T25 did not produce mRNA from the disrupted <i>bla</i> gene, nor did they produce beta-lactamase, the enzyme encoded by the <i>bla</i> gene. Additionally, the <i>bla</i> gene would not have been expected to be functional in the modified maize lines, as it did not have the necessary regulatory sequences for expression in plants.</p>

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

<p style="text-align: justify;">The integration of the transferred DNA, into events T14 and T25, was demonstrated to be stable by segregation analysis. The data provided demonstrated the integration of one and three copies of the <i>pat</i> gene in transformation events T25 and T14, respectively. Commercial maize lines containing the transformation events are several generations removed from the two original transformation events, further demonstrating the stable inheritance of the herbicide resistant trait.</p>

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

<p style="text-align: justify;">The only newly expressed protein in T14- and T25-derived plants was the PAT protein. The <i>pat</i> gene was linked to a constitutive promoter, and protein expression was detected in plant tissues, grain and silage. Expression of the <i>pat</i> gene in T14 and T25 resulted in the production of PAT protein in maize tissues at low levels (&lt; 0.003% (w/w) of total crude protein). The PAT enzyme was not detected in protein extracts from the pollen.</p>
