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

<p style="text-align: justify;">Southern blot analysis of MON 88913 genomic DNA indicated the incorporation of a single intact integration of the T-DNA insert of PV-GHGT35, at a single locus. The analysis revealed the intact insertion of the two <i>cp4 epsps</i> cassettes, including the promoter, enhancer, terminator, and chlorophyll transit peptide (<i>ctp</i>2) sequences. Evidence was also provided to show that none of the plasmid backbone sequences were incorporated into the genome of MON 88913 cotton.</p>

<p style="text-align: justify;">The <i>cp4 epsps</i> gene was isolated from <i>Agrobacterium tumefaciens</i> strain CP4 and encodes a 47.2 kDa EPSPS protein. In plants, EPSPS is a chloroplast-localized enzyme that is transported from the cytosol to the chloroplast by the chlorophyll transit peptide. In MON 88913, the inserted <i>ctp</i>2 gene codes for a chlorophyll transit peptide that binds to the CP4 EPSPS enzyme to enable transportation to the chloroplast.</p>

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

<p style="text-align: justify;">The stability of the glyphosate tolerance trait in MON 88913 was assessed within and across generations using Southern blot analysis. Progeny resulting from the self-pollination of first generation plants segregated in a 3:1 of resistant to non-resistant plants. Advanced generations (R4 and R5) of self-pollinated homozygous plants resulted in segregation ratios of 1:0. These results were not significantly different from the expected Mendelian segregation ratios and confirmed the homozygosity and generation stability of a single dominant trait across multiple generations.</p>

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

<p style="text-align: justify;">The levels of CP4 EPSPS in tissues of MON 88913 were quantified using enzyme-linked immunosorbant assay (ELISA) methods. Samples for analysis were obtained from field trials at four different locations in 2002. Levels of CP4 EPSPS expression were determined on a fresh weight basis, and also calculated on a dry weight basis from dry matter percentages determined for the various tissues. Dry matter percentage was not determined for pollen due to the limited quantifies available for analysis. The average expression of CP4 EPSPS protein, on a fresh weight (dry weight) basis, was: 170 (970) &micro;g/g in young leaves, 160 to 270 (630 to 1400) &micro;g/g in overseason leaves, 31 (99) &micro;g/g in roots, 310 (340) &micro;g/g in seeds, and 4 &micro;g/g (fresh weight only) in pollen. The levels of expression of CP4 EPSPS are higher in MON 88913, compared to the previously approved Roundup Ready Cotton.</p>
