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

<p style="text-align: justify;">The <i>cry9C</i> gene was synthesized with plant preferred codons before it was stably inserted into maize plants to produce a truncated and modified Cry9C protein. The CBH-351 expressed Cry9C protein was similar to the trypsin-resistant core protein produced by B. thuringiensis, with the exception of a single amino acid substitution in the internal sequence and the addition of two extra amino acids to the N-terminus.</p>

<p style="text-align: justify;">The modifications that have been introduced into the Cry9C protein expressed in CBH-351 maize, as compared to the wild type 129.8 kDa Cry9C protoxin include the following:</p>

<ol>
	<li style="text-align: justify;">C-terminal truncation that removes all the amino acids (aa) following position 666 of the wild type protoxin just after the conserved sequence block 5 shared by certain Lepidoperta-active Bt Cry proteins;</li>
	<li style="text-align: justify;">&nbsp;N-terminal truncation that removes the first 43 aa at the point corresponding to the N-terminus of the 68.7 kDa fragment of the wild type toxin produced by initial trypsin-digestion, and the subsequent N-terminal addition of the amino acids methionine and alanine;</li>
	<li style="text-align: justify;">Replacement of arginine by lysine at position 123 in the plant encoded protein which reduces the susceptibility of the protein to trypsin cleavage to a non-toxic 55 kDa fragment.</li>
</ol>

<p style="text-align: justify;">&nbsp;</p>

<p style="text-align: justify;">These modifications did not appear to affect the insecticidal activity of the Cry9C protein against the primary targeted insect, European corn borer (ECB), <i>Ostrinia nubilalis</i>.</p>

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

<p style="text-align: justify;">Molecular and biochemical analyses presented for maize line CBH-351 demonstrated that the number of insertion sites, copy number, and expression level were stably inherited.</p>
