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

<p style="text-align: justify;">Plasmid PV-ZMPQ76 contained three expression cassettes &ndash; <i>cordapA</i>, <i>nptII</i>, and <i>amp</i> which was used for selection in <i>E.coli</i>. The 5.9 Kb fragment of this plasmid used for transformation contained only the <i>cordapA</i> and <i>nptII</i> cassettes and in the final product only the <i>cordapA</i> cassette was present. This was confirmed by event-specific PCR analyses and Southern blot analyses, which also confirmed that LY038 contained one intact copy of the <i>cordapA</i> gene cassette. No additional elements from PV-ZMPQ76 were identified in the genome and there were no intact or partial DNA fragments from PV-ZM003 (used to transform lines expressing the Cre recombinase).</p>

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

<p style="text-align: justify;">Heritability and stability of the <i>cordapA</i> gene cassette in LY038 were determined on plants from: 1) the F1&#39; generation (prior to excision of the <i>npt II</i> marker gene); 2) the F3 generation (after the excision of the <i>npt II</i> marker gene); and 3) the F4 generation (after two rounds of backcrossing to conventional inbred lines). The expected segregation ratios were 1:1, 3:1, and 3:1, for the F1&#39;, F3, and F4 generations, respectively. There were no significant differences in the observed-to-expected segregation ratios for the LY038 <i>cordapA</i> gene cassette over five plant generations, as demonstrated by the chi-square values. These segregation data indicates a single-locus Mendelian inheritance pattern for the transgene. These data are also consistent with the molecular analyses which suggested that the <i>cordapA</i> transgene was stably integrated at a single site in the genome. Southern blot analysis of LY038 genomic DNA established that the inserted DNA was stably transferred across seven generations and that <i>cre</i> and <i>nptII</i> cassette elements were absent in LY038.</p>
