This paper reports the isolation and characterization of a complementary deoxyribonucleic acid clone showing sequence homology with genes coding for the eukaryotic elongation factor 1 gamma (EF-1 gamma). The clone encodes an open reading frame of 404 amino acids corresponding to a deduced molecular mass of 46.2 kDa. Database searches revealed 30-64% sequence identity between the Leishmania infantum EF-1 gamma and several eukaryotic homologues. Southern blot analysis indicated that 2 genes tandemly organized were present in the L. infantum genome. The 3' untranslated regions of these 2 genes differed in size. Southern hybridization and pulsed field gel electrophoresis showed that EF-1 gamma genes are highly conserved among members of the Leishmania genus and must be clustered in a single chromosomal locus.