Cry3A gene obtained from Bacillus thuringiensis. The gene was modified for enhanced expression in maize and such that the amino acid sequence of the synthetic version of Cry3A is the same as the native protein, except for the modified serine-protease recognition site.

The pmi gene encodes the enzyme phosphomannose isomerase (PMI) that allows the plants to utilise mannose as a carbon source and is used as a selectable marker.

Truncations at the left and right border junctions of the T-DNA insert as well as nucleotide changes were identified relative to the intended DNA sequence. These substitutions have not resulted in any apparent functional change in PMI as expressed in MIR604.

Southern hybridization data provide confirmatory evidence to support the Taqman PCR analysis that Corn MIR604 contains a single copy of mcry3A gene and the pmi gene.

Corn MIR604 contains a single copy of the MTL and ZmUbilnt promoters, without any vector backbone sequences present in pZM26. Sequence analysis revealed that truncations and deletions occurred but have no effect on the efficacy of the T-DNA insert. Three base pair changes were noted; one occurred in the regulatory region and does not encode for a protein; two other base pair changes occurred within the coding region but these amino acid changes did not result in any apparent functional change in the new insert.

As expected for the mcry3A, pmi, MTL and ZmUbilnt probes the Kpn1 digest resulted in a single hybridization band demonstrating that a single copy of each element is present in Corn MIR604.  Additionally, for the full length backbone probe, lack of hybridization demonstrates the absence of any pZM26 vector backbone sequences being incorporated into corn MIR604 during the transformation process.