To understand molecular mechanism of spermatogenesis, two ESTs BG720564 and AI700454, were found from SPAG4 (sperm antigen 4), a gene related to dense fiber protein of outer membrane of the human sperm and mouse spermatocytes gene AK006225. The gap was filled by polymerase chain reaction, and a 1252 bp fragment was obtained. This 1252 bp fragment was named TSARG4 (testis and spermatogenesis related gene 4 (GenBank accession number AF401350). Its opening reading frame was 94-1233 bp, as was proved by RT-PCR in human testis. TSARG4 gene was located in 20q11.2, and the putative protein was 379 amino acid with a theoretical molecular weight of 43 081 and isoelectric point of 8.61. The homologies of amino acid sequences were 74% between TSARG4 and AK006225 gene and 45% between TSARG4 and SPAG4 gene, respectively. The human TSARG4 mRNA is expressed in a wide range of adult tissues, including testis, whereas the homologous mouse spermatocytes gene AK006225 is expressed specifically in the testis.