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

<p style="text-align: justify;"><u>Line RBMT15-101</u> - insertion of the T-DNA occurred at three to four loci. At least one locus contained two copies of the T-DNA organized in inverted orientations. For two copies of the T-DNA, transfer was incomplete at the right border resulting in an incomplete copy of the figwort mosaic virus (FMV) 35S promoter associated with the PVY CP gene. One of the <i>cry3Aa</i> genes also lacked the Arabidopsis small subunit promoter and a portion of the 5&#39; end of the gene. The NOS terminator region of this gene cassette was intact. One of the T-DNAs also had an incomplete NOS promoter region associated with an intact NPTII coding region. The coding regions of all the other genetic elements were intact. The analyses also showed that no plasmid sequences beyond the left and right borders were transferred.</p>

<p style="text-align: justify;"><u>Line SEMT15-02</u> - insertion of the T-DNA occurred at four to five loci. At least one locus contained two copies of the T-DNA organized in inverted orientations and one locus contained two T-DNAs linked by a complete copy of the plasmid backbone. For seven copies of the T-DNA, transfer of the T-DNA resulted in incomplete resolution of the right border, leaving incomplete copies of the FMV promoter associated with the PVY CP coding region. Plasmid sequences beyond the left and right borders, which include the <i>aad</i> gene and the oriV and ori322 plasmid elements, were inserted into this line. Integration of complete backbone elements occurred in two different ways: at one locus two T-DNAs were linked by a complete copy of the backbone; at two other loci, backbone integration was not associated with the left border flanking the NOS promoter of the NPTII encoding gene.</p>

<p style="text-align: justify;"><u>Line SEMT15-15</u> - insertion of the T-DNA occurred at four to five loci. At least one locus contained copies of the T-DNA organized in inverted orientations. For two copies of the T-DNA, transfer of the T-DNA resulted in incomplete resolution of the right border, leaving incomplete copies of the FMV promoter associated with the PVY CP coding region. One of the T-DNAs in this line had an incomplete NOS promoter region associated with an intact NPTII coding region. The coding regions of all the genetic elements were intact. Plasmid sequences beyond the left and right borders, which include the <i>aad</i> gene and the oriV and ori322 plasmid elements, were inserted into this line.</p>

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

<p style="text-align: justify;">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 inframe 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 RBMT15-101, SEMT15-02 and SEMT15-15 potatoes.</p>

<p style="text-align: justify;">The levels of expressed Cry3A protein were quantitated using enzyme linked immunosorbent assay (ELISA) of samples of leaf tissue from field grown plants and were found to range between 20-63 &micro;g/g fresh weight tissue. Concentrations of Cry3A in tuber tissue (0.08 - 0.38 &micro;g/g) were approximately 100 fold lower than that present in leaf tissue, equivalent to 0.0005 - 0.0019% of total tuber protein.</p>

<p style="text-align: justify;">The transgenic expression of PVY CP was estimated to be less than the detectable limit of 2 ng/mg fresh leaf tissue. In contrast, the accumulation of PVY-O coat protein in naturally virus-infected potato plants was approximately 12 to 244-fold greater, in the range of 24-488 ng/mg fresh leaf tissue when measured by ELISA. NPTII was expressed in the tuber at levels, ranging from 0.003 - 0.01 &micro;g/g fresh weight (equivalent to &lt; 0.001% of the total tuber protein).</p>
