<p style="text-align: justify;">The soybean sublines G94-1, G94-19 and G168 were produced via biolistic transformation of the parent soybean line (Asgrow A2396) with a mixture of two plasmids pBS43 and pML102. These sublines were advanced using traditional breeding techniques to produce high oleic soybean varieties.</p>

<p style="text-align: justify;">Plasmid pBS43 contained the GmFad2-1 gene of soybean (<i>Glycine max</i>) and the beta-glucuronidase (GUS; <i>uidA</i>) gene from <i>Escherichia coli</i>. The GmFad2-1 gene was inserted in the sense orientation and expression of the GmFad2-1 was regulated by a seed specific promoter from the soybean beta-conglycinin gene and a transcription terminator from the phaseolin gene from <i>Phaseolus vulgaris</i>. The <i>uidA</i> gene was used during the transformation process as a reporter gene to select plants cells which carried the introduced genes. Expression of the GUS encoding gene was controlled by the 35S promoter from cauliflower mosaic virus (CaMV) and a nopaline synthase (nos3&#39;) terminator.</p>

<p style="text-align: justify;">Plasmid pML102 contained the <i>dapA</i> gene from <i>Corynebacterium glutamicum</i> which encodes the enzyme dihydrodipicolinic acid synthase (DHDPS). Corynebacterium DHDPS catalyzes a step in the biosynthesis of the amino acid lysine but is insensitive to feedback inhibition by lysine. Expression of this enzyme in soybean seeds results in accumulation of the essential amino acid lysine. The <i>dapA</i> gene is controlled by the seed specific promoter Kunitz trypsin inhibitor (Kti3) from soybean that allows high levels of expression. The <i>dapA</i> gene was fused to a chloroplast transit peptide sequence to direct the DHDPS protein into the chloroplast where lysine biosynthesis is carried out.</p>

<p style="text-align: justify;">Both plasmids contained the antibiotic resistance marker, beta-lactamase (<i>bla</i>) gene. The <i>bla</i> gene confers resistance to the antibiotic ampicillin and permits the selection of transformed <i>E. coli</i> cells during laboratory recombinant DNA steps. The <i>bla</i> gene contains its own <i>E. coli</i> regulatory sequences and was therefore not expressed in the transformed plants.</p>
