<p style="text-align: justify;">Maize line MON 863 was produced by biolistic transformation of the inbred line A634 (reportedly one of the most popular publicly available inbred lines used in hybrid maize production in the United States) using linearized plasmid PV-ZMIR13 DNA purified following <i>Mlu I</i> restriction endonuclease digestion. The introduced DNA contained the modified <i>cry3Bb1</i> gene from <i>B. thuringiensis</i> subsp. <i>kumamotoensis</i> under the control of the 4-AS1 promoter (CaMV 35S promoter with 4 repeats of an activating sequence), plus the 5&#39; untranslated leader sequence of the wheat chlorophyll a/b binding protein (wt CAB leader) and the rice actin intron. The transcription termination sequence was provided from the 3&#39; untranslated region of the wheat 17.3 kD heat shock protein (<i>tahsp17</i>). The modified <i>cry3Bb1</i> gene encodes a protein of 653 amino acids whose amino acid sequence differs from that of the wild-type protein by the addition of an alanine residue at position 2 and by seven amino acid changes.</p>

<p style="text-align: justify;">The introduced DNA also contained a copy of the neomycin phosphotransferase II (NPTII) encoding gene (<i>neo</i>; synonym <i>nptII</i>) derived from the Tn5 transposon of <i>Escherichia coli</i>. This gene was introduced as a selectable marker under the control of the CaMV 35S promoter and the 3&rsquo; untranslated termination sequence (NOS 3&rsquo;) from the <i>Agrobacterium tumefaciens</i> nopaline synthase gene. The <i>nptII</i> cassette also contained a 153 base pair portion of the bleomycin binding protein gene (<i>ble</i>; 378 bp total size).</p>
