A recent genetic study in humans where the ZFPM2 gene was screened for mutations revealed a heterozygous mutation that was found to contribute to several sporadic cases of tetralogy of Fallot (TOF). We hypothesized that this would also hold true for a study group of dogs with TOF. We isolated the full-length cDNA of the canine ZFPM2 and aligned the deduced amino acid sequence against sequences from other species. Our analysis indicated that the canine ZFPM2 was phylogenetically much closer to the human ZFPM2 compared with the mouse or rat sequences. Further gene expression analysis also failed to identify differential gene expression between affected and unaffected dogs. We screened the cZFPM2 for mutations in a group of dogs with TOF but found no significant base changes in either the splice sites or the coding regions.