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

<p style="text-align: justify;">Southern blot experiments confirmed the presence of a single copy the <i>cry1ab</i> and <i>pat</i> genes in BT11 maize lines and the absence of the beta-lactamase encoding <i>bla</i> gene.</p>

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

<p style="text-align: justify;">The stability of the inserted DNA in BT11 maize was demonstrated by a Mendelian inheritance pattern using Southern blot analysis. Segregation analysis of the <i>cry1Ab</i> and <i>pat</i> genes over multiple generations demonstrated that they were closely linked, as they always segregated together. Restriction fragment length polymorphism (RFLP) mapping was used to determine the location of the novel genes in BT11. The single insertion site was mapped to the long arm of chromosome 8.</p>

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

<p style="text-align: justify;">The expression levels of both the Cry1Ab protein and the PAT enzyme were determined in the leaves and kernels of transgenic corn. Accounting for extraction efficiencies, the amount of expressed Cry1Ab protein was found to range between 15 &ndash; 29 &micro;g/g fresh weight for mature and young leaves, respectively, and 3.7 or 4.76 &micro;g/g fresh tissue for heterozygous or homozygous kernels. The PAT enzyme was detected at levels of 38.6 &ndash; 49.4 ng/g fresh weight of leaf tissue, but not in roots, pollen or kernels. However, enzyme activity analysis indicated that PAT was expressed in all tissues throughout the plant. No significant differences were observed between hybrids derived using original elite lines and the selected BT11 line for the agronomic traits of yield, moisture at harvest, root lodging rating, ear height, plant height, heat units to silking or pollen shed. Other than resistance to ECB and tolerance to glufosinate ammonium herbicide, the disease, pest and other agronomic characteristics of BT11 corn were comparable to non-transgenic lines of corn.</p>
