<p style="text-align: justify;">MON 88913 cotton was produced by <i>Agrobacterium</i>-mediated transformation of hypocotyl tissue from the cotton variety &lsquo;Coker 312.&rsquo; The binary plasmid PV-GHGT35 was used for the transformation. The T-DNA portion of the plasmid contained two tandem <i>cp4 epsps</i> gene expression cassettes. Joined with each <i>cp4 epsps</i> gene was a chloroplast transit peptide (<i>cpt</i>2) sequence, derived from the <i>Arabidopsis thaliana epsps</i> gene. Two unique promoter and enhancer sequences were used to regulate the expression of each <i>cp4 epsps</i> gene. In the first cassette, gene expression was regulated by P-FMV/TSF1, a chimeric promoter containing the <i>A. thaliana tsf1</i> gene promoter, the encoding elongation factor EF-1 alpha, enhancer sequences from the Figwort Mosaic Virus 35S promoter, as well as the leader and intron sequences from the <i>A. thaliana tsf1</i> gene. The expression of the second <i>cp4 epsps</i> gene was regulated by P-35S/ACT8, a chimeric promoter from the <i>act8</i> gene of <i>A. thaliana</i> combined with enhancer sequences from the Cauliflower Mosaic Virus 35S promoter, and the leader and intron sequences from the <i>act8</i> gene of <i>A. thaliana</i>. The transcriptional termination and polyadenylation sequences were derived from the T-E9 DNA sequences of <i>Pisum sativum</i>, containing the 3&rsquo; nontranslated region of the ribulose-1,5-biphosphate carboxylase (Rubisco) small subunit E9 gene.</p>

<p style="text-align: justify;">Transformed callus was selected <i>in vitro</i> using culture medium to which glyphosate had been incorporated. Plants were regenerated from tolerant callus using tissue culture techniques. MON 88913 was derived from a single regenerated plant that displayed tolerance to glyphosate.</p>
