Access to the nucleotide sequence of parvalbumin from chicken muscle was gained via the polymerase chain reaction. In the absence of specific amino acid sequence data, the PCR primers were based on consensus data for the two parvalbumin Ca2(+)-binding sites. The 137 bp fragment obtained by amplification clearly codes for a parvalbumin, as judged by the presence of 10 invariant codons within the sequence flanked by the primers. When used to probe poly(A)+ RNA from chicken muscle, the fragment recognizes an 800 nucleotide transcript. The translated nucleotide sequence of the muscle protein is unmistakably distinct from that of the thymus-specific parvalbumin known as avian thymic hormone. Of the 31 amplified residues, the two proteins differ at 14. The presence of a distinct parvalbumin in chicken thymus is consistent with the potent effector role proposed for the protein.