Since the plasmid form of the AquAdvantage DNA construct,
opAFP-GHc2, comprises the complementary deoxyribonucleic acid
(cDNA) sequence of a Chinook salmon GH1 gene as an integrated
transcriptional unit, sequence comparison of the Chinook (C) and
Atlantic (A) salmon GH1 genes was performed in order to ensure
the non-occurrence of cross contaminations. For this, a protein
and nucleotide blast was studied with the sequences of salmon
GH1. Growth hormone gene 1 (GH1) from salmon fish species
was selected for setup using the qPCR method. Complete and partial
sequences from different genera and species of the Salmonidae
family were retrieved from the GenBank database (Data not
shown). The web multialign program (http://multalin.toulouse.
inra.fr/) was used for sequence alignments. The study of theoretical
specificity corresponded to the choice of a region that makes the
difference between Atlantic salmon (S. salar L.) and all other species
of salmonids. The alignment allowed us to choose the adequate
positions of the primers and probe for the differentiation
between the trout (Salmo trutta) and the salmon S. salar GH1 and
thus avoiding false positive results in further amplifications with
real-time PCR. The results of the sequence alignment and the primer
and probe locations are shown in