To facilitate the characterization of genetic expression in human adult testis, expressed sequence tag analysis of cDNAs from this tissue has been undertaken. Over 180 kb of DNA sequence has been determined and used to search the GenBank database. The results from the first 359 cDNA clones analyzed indicate that the sequences could be sorted into several categories with a high proportion being novel. Twenty-five clones (7%) showed 100% identity with human genes, 11 (3%) with prokaryotic sequences, 21 (5%) with between 60 and 95% similarity to human genes, 27 (8%) with between 60 and 95% similarity to genes from other species, and 33 (9%) with matches to human repeat sequences. Two hundred forty-two (67%) showed no significant matches and thus are likely to represent novel transcripts. In comparison to similar studies on human brain tissue and a hepatoma cell line, the findings indicate that the matches in the testis transcript population appear to be identifying a different spectrum of gene sequences. (C) 1994 Academic Press, Inc.