<p style="text-align: justify;">The transgenic tomato line 5345 was produced via <i>Agrobacterium</i>-mediated transformation of UC82B tomato using a protocol in which the transfer-DNA (T-DNA) of the bacterial tumour inducing (Ti) plasmid was modified to contain genes encoding the Cry1Ac delta-endotoxin from <i>B. thuringiensis</i> subsp kurstaki, NPTII, and another enzyme used as a bacterial selectable marker, 3&quot;(9)-O-aminoglycoside adenylyltransferase (AAD). The AAD encoding gene, which was not expressed in the plants, was used during the development process to select for bacterial colonies that had been transformed with recombinant plasmid DNA.</p>

<p style="text-align: justify;">The nucleotide sequence of the <i>cry1Ac</i> gene was modified via site-directed mutagenesis to contain plant-preferred codons in order to maximize protein expression in plant cells. These modifications did not alter the predicted amino acid sequence of the Cry1Ac protein. Expression of the introduced <i>cry1Ac</i> gene was regulated by including promoter sequences derived from the 35S transcript of cauliflower mosaic virus (CaMV) and a 3&rsquo; non-translated region of the soybean alpha subunit of the beta-conglycinin gene. The <i>neo</i> gene was regulated using the <i>nos</i> promoter and terminator sequences from <i>A. tumefaciens</i>.</p>
