Mx proteins are GTPases that are specifically induced by type I interferons (IFN) in vertebrates. Some mammalian Mx proteins have antiviral activity against certain RNA viruses. A 2.3-kb full-length cDNA clone of an Atlantic halibut Mx gene was isolated from a liver cDNA library. The open reading frame (ORF) predicts a 622 amino acid protein of 71.2 kDa possessing a tripartite GTP binding motif, a dynamin signature, and a leucine zipper motif, which are conserved in all known Mx proteins. The C-terminal half contains a putative bipartite nuclear localization signal. The deduced halibut Mx protein showed approximately 76% sequence identity with the Atlantic salmon and rainbow trout Mx proteins, 55% identity with the human MxA, and 48% identity with the chicken Mx protein. Based on sequence comparison of 554-bp Mx cDNA fragments, the Atlantic halibut Mx showed more relationship with the perch and turbot than the salmonid Mx genes. Halibut appears to possess at least two Mx loci, as suggested by Southern blot analysis of genomic DNA. Two halibut Mx transcripts (2.2 kb and 2.6 kb) were strongly induced in vivo by the double-stranded RNA (dsRNA) poly I:C or infectious pancreatic necrosis virus (IPNV) in all organs studied.