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

<p style="text-align: justify;">The data provided showed various integration patterns depending on the line: the T- DNA was integrated at three sites for line ATBT04-6 and at one site for lines ATBT04-30 and ATBT04-31. No coding region from outside the T-DNA borders was incorporated into the genotype of these lines. One complete copy of the T-DNA was found at each of two insertion sites in line ATBT04-27, with one site also containing a bacterial gene coding for resistance to streptomycin and another partial copy of the <i>cry3A</i> gene. The streptomycin resistance gene is driven by a bacterial promoter and is not expressed in plants, as was demonstrated in these NewLeaf&trade; Atlantic potatoes. Finally, the transgenes were integrated at three insertion sites in line ATBT04-36.</p>

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

<p style="text-align: justify;">The NewLeaf&trade; potato lines were evaluated over several generations of vegetative propagation, used to generate potato seed, to determine if they expressed consistent levels of the Cry3A protein in leaf tissue. Expression of the Cry3A protein was found to be highly stable across multiple generations in all the lines tested.</p>

<p style="text-align: justify;"><b>Expressed Material</b><br />
In <i>B. thuringiensis</i>, expression of the <i>cry3A</i> gene results in the production of two proteins, a full length 73 kDa (644 amino acids) Cry3A protein and a smaller molecular weight species (68 kDa; 597 amino acids) resulting from in-frame translation initiation at an internal ATG sequence. This latter protein, referred to as Cry3A band 3 protein, lacks 48 amino acids from the N-terminus but retains insecticidal activity, and corresponds to the form of the protein expressed in these transgenic potatoes.</p>

<p style="text-align: justify;">Western immunoblot analysis of protein extracts from transgenic potato tissue demonstrated the presence of a 68 kDa immunoreactive species as well as a smaller molecular weight species of 55 kDa. Studies were conducted to confirm that the 68 kDa protein product of the <i>cry3A</i> gene transferred into potato plants undergoes processing or degradation in the plant cell into a smaller protein of 55 kDa. The two protein products seen in the plant cell appear identical to that observed in commercial B.t.t preparations.</p>

<p style="text-align: justify;">Protein expression levels were quantitated by enzyme linked immunosorbent assay (ELISA) and found to average from 15.7 to 59.3 &micro;g/g (f.w.) of leaf tissue and from 0.09 to 0.53 &micro;g/g (f.w.) of tuber tissue, corresponding to 0.10 to 0.37% of total foliage protein and 0.0004 to 0.003% of total tuber protein. Likewise, production of NPTII was quantified in both leaf and tuber tissue at levels of 4.4-36.6 and 0.5-2.9 &micro;g/g fresh weight tissue, respectively. The presence of NPTII protein has been judged to be insignificant with respect to any human health risk due to exposure.</p>
