DNA fragment No. 13 from the C. psittaci pigeon strain, has been cloned in the plasmid pUC19. Hybridization analysis revealed that the fragment maintained a chlamydial common sequence. Furthermore, nucleotide sequencing identified two partial open reading frames (ORF), 675b. p. and 530b. p.. Expression of ORFs revealed that the second ORF encoded 25KD polypeptide, whereas the first ORF did not produce any antigenic product. The 25KD beta-galactosidase fusion protein reacted strongly with chlamydia-specific antibodies elicited against a number of different chlamydial strains. Gene Bank analysis showed that this cloned gene is not highly homologous with chlamydia or other organisms for which nucleotide sequences have already been published. This 25KD polypeptide may be an additional genus-specific antigen of Chlamydiae.