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

<p style="text-align: justify;">Analysis of the DNA introduced into the DBT418 genome indicated integration of two intact copies of the <i>cry1Ac</i> gene, one intact copy of the <i>bar</i> gene and one rearranged copy of the <i>bar</i> gene. The incorporation of the <i>pinII</i> gene was incomplete due to rearrangements during or following the transformation process. There are four intact and one partial copy of the beta-lactamase (<i>bla</i>) gene and four intact copies of the <i>ColE1</i> origin of replication. The beta lactamase gene was not expressed and the <i>pinII</i> gene was non-functional and therefore not expressed.</p>

<p style="text-align: justify;">The synthetic <i>cry1Ac</i> gene introduced into line DBT418 encoded the N-terminal 613 amino acids of the native Cry1Ac protein and was modified to reflect plant preferred codon frequencies in order to maximize translation in maize cells. The gene codes for a 66 kDa protein of similar molecular weight to the protein tryptic core fragment of the <i>Bacillus thuringiensis</i> var. kurstaki HD-73 insecticidal crystal Cry1Ac protein.</p>

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

<p style="text-align: justify;">The expression of Cry1Ac and PAT proteins was demonstrated to be consistent across several generations in several different genetic backgrounds.</p>

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

<p style="text-align: justify;">Expression of the Cry1Ac protein and PAT occured in most, but not all, tissues of the maize plant with levels highest in leaves, and lower levels in roots, prop roots, stalk, tassel, cob, husk, and kernels. No Cry1Ac or PAT proteins were detected in silk or pollen.</p>

<p style="text-align: justify;">Average protein expression of Cry1Ac in various tissues were as follows: harvested leaves ranged from 459.6 to 1194.4 ng Cry1Ac per gram tissue (dry weight); harvested kernels ranged from 36.0 to 42.8 ng Cry1Ac per gram tissue (d.w); harvested roots ranged from 58.0 to 125.4 ng Cry1Ac per gram tissue (d.w.); harvested stalks ranged from 40.9 to 123.6 ng Cry1Ac per gram tissue (d.w.).</p>

<p style="text-align: justify;">The expressed PAT enzyme was compared to the bacterial protein: molecular weights were similar, indicating that the protein had not been glycosylated nor had it undergone post transcriptional modifications. The amino terminal sequence of PAT derived from DBT418 was determined to be the same as the native protein.</p>

<p style="text-align: justify;">Average protein expression of PAT in various tissues, sampled at the pollen stage (except kernels), were as follows: leaves ranged from 501.8 to 1099.4 &micro;g PAT per g tissue (d.w.); stalks ranged from 66 to 77 &micro;g PAT per g tissue (d.w.),roots ranged from 27.5 to 69.5 &micro;g PAT per g tissue (d.w.), and kernels ranged from 3.1 to 6.0 &micro;g PAT per g tissue (d.w.).</p>
