Rat SPATA4 gene, homologue to the human and mouse SPATA4 gene, expressed specifically in the rat testis was cloned by informatics analysis. The cDNA mapped to chromosome 16 in the rat genome is made up of 6 exons and the exon-intron boundaries obey to the AG/GT rule. The gene contains a 972 bp open reading frame encoding 323 amino acid sequences with theoretical molecular weight of 36.64 KD and isoelectric point of 9.65. One CpG island is located in the gene from site -200 to +198. A typical promoter is also predicted from site -630 to +101. According to the computer-aided analysis of the putative protein encoded by the rat SPATA4, no transmembrane region and no signal peptides are found in the protein. Multi-tissue RT-PCR results show that the SPATA4 gene is expressed specifically in the testis only. Moreover, the expression of SPATA4 occurs in a development stage-dependent pattern. According to the RT-PCR results, no expression of SPATA4 is detected until the rat is 30 d old after birth. The amount of SPATA4 mRNA increases from 30-d to 65-d-old rat and then keeps stable after that. In conclusion, this study proves the conservation of SPATA4 in mammalian animals and predicts its important role in spermatogenesis.