The applicant showed by Southern blot analyses that one copy of the insert with the bar and cry2Ae gene is integrated at a single locus. The applicant further showed that the backbone of the plasmid used for transformation is absent in GHB119. Bioinformatic analysis indicated that Cry2Ae and PAT protein in GHB119 cotton has no homology to any known toxins, using FASTA algorithm and BLOSUM50 scoring matrix.

In relation to potential toxicity and allergenicity, it is worth noting that Cry2Ae and PAT proteins are inherently non-toxic to mammals and do not exhibit any potential to be allergenic to humans. In addition, bioinformatic studies have confirmed their lack of any significant amino acid sequence similarity to known protein toxins or allergens and digestibility studies have demonstrated that both proteins would be rapidly degraded in the stomach following ingestion. Acute oral toxicity studies in mice have also confirmed their absence of toxicity in animals. Both proteins exhibit a degree of heat stability, however given their digestive lability, this does not raise any safety concerns. Taken together, the evidence indicates that Cry2Ae and PAT are unlikely to be toxic or allergenic to humans.

In terms of the interaction among the two novel proteins, no likelihood of interaction among the novel proteins, Cry2Ae and PAT since they are expressed independently of each other and their mode of actions and metabolic pathways are different from one another.