The ID (inhibitors of DNA binding/differentiation) proteins represent a family of dominant negative regulators of the basic helix-loop-helix transcription factors whose activities result in delayed cell differentiation and prolonged proliferation. A pair of expressed sequence tag clones with homologies to the ID proteins were identified and used to screen a rainbow trout bacterial artificial chromosome (BAC) library to identify clones containing homologues sequences. Phylogenetic analysis of the predicted amino acid sequences revealed close similarities to the rainbow trout ID1 protein, the genes were therefore classified as rainbow trout ID1B and ID1C. Genome characterization based on BAC sequencing showed each gene to have two exons separated by a small intron. The genes are 83% similar in their transcribed regions, yet they are only 64 and 65% similar in the upstream and downstream sequences, respectively. Using reverse transcription polymerase chain reaction, we found both genes to be expressed in a variety of tissues in the adult rainbow trout.