The nucleotide sequence of the Clostridium perfringens chloramphenicol acetyltransferase (CAT)-encoding resistance determinant, catQ, was determined. An open reading frame encoding a protein of 219 amino acids with a molecular weight of 26,014 was identified. Although catQ was expressed constitutively, sequences similar in structure to those found upstream of inducible cat genes were observed. The catQ gene was distinct from the C. perfringens catP determinant. The deduced CATQ monomer had considerable amino acid sequence conservation compared with CATP (53% similarity) and other known CAT proteins (39 to 53%). Phylogenetic analysis revealed that the CATQ monomer was as closely related to CAT proteins from Staphylococcus aureus and Campylobacter coli as it was to CAT monomers from the clostridia.