A cDNA clone encoding ribosomal protein S27 (AmphiS27) was identified in the gut cDNA library of amphioxus Branchiostoma belcheri tsingtauense. This cDNA consists of 607 bp and contains a 255 bp open reading frame (ORF) corresponding to a deduced protein of 84 amino acids with a calculated molecular mass of 9,488 Da and an isoelectric point (pI) of 9.500. Alignment of the deduced AmphiS27 amino acid sequence with 12 known S27 protein sequences indicates that AmphiS27 shares 94-99% homology with its vertebrate homologue, 84-94% with invertebrate homologues and 69-72% with homologues from other eukaryotes, suggesting that AmphiS27 is more closely related to the vertebrate S27 protein than to its invertebrate counterpart. Southern blot analysis showed a single copy of the S27 gene present in the genome of amphioxus B. belcheri tsingtauense, indicating that amphioxus has a genome uncomplicated by extensive gene duplication.