The NLRs (nucleotide-binding domain and leucine-rich repeat containing family receptors) are a recently identified family of pattern recognition receptors in vertebrates. Several subfamilies of NLRs have been characterized in human, mouse, and zebrafish, but studies of NLRs in other species, especially teleost species, have been lacking. Here we report characterization of five NLRs from channel catfish: NOD1, NOD2, NLRC3, NLRC5, and NLRX1. Structural analysis indicated that the genes were organized in a similar fashion as in the mammals and in zebrafish. Phylogenetic analysis suggested that they were orthologous to the NOD-like subfamily of NLRs. All five NOD-like genes exist as a single copy gene in the catfish genome. Hybridization of gene-specific probes allowed mapping of three NLR genes to the catfish physical map, laying a foundation for genome characterization and for establishing orthologies with NLR genes from other species. These genes are widely expressed in various tissues and leukocyte cell lines. While the majority of the NLR genes appeared to be constitutively expressed, NOD1 was induced after infection with a bacterial pathogen, Edwardsiella ictaluri, the causative agent of enteric septicemia of catfish (ESC), suggesting its involvement in immunity against the intracellular pathogen.