In order to isolate, clone, and sequence agouti exon 2 of the pig (Yorkshire), we used an interspecific hybridization strategy. Primers from the 5′ and 3′ borders of the known human agouti exon 2 sequence were used to amplify (PCR) pig agouti exon 2. Following Southern blotting using a human exon 2 internal primer to authenticate that our PCR amplified product was truly pig exon 2 (PorAex2), the fragment was cloned and sequenced. PorAex2 exhibits 79.1 and 75.7% DNA sequence and 85 and 74% deduced amino acid sequence homologies with human and mouse agouti exon 2 and agouti protein, respectively. With the isolation of PorAex2, we can now map, sequence, and clarify the modus operandi of the porcine agouti gene. The GenBank Accession number of PorAex 2 is AF018166.