Vasa gene homologs of tiger puffer (Takifugu rubripes, Tp‒vasa) and grass puffer (T. niphobles, Gp‒vasa) were cloned and characterized for use as a molecular marker of germ cells in these species. Analysis of the nucleotide sequence revealed that open reading frames of Tp‒ and GP‒vasa comprise 1,872bp (encoding 623 amino acids) and 1,869bp (encoding 622 amino acids), respectively. These deduced amino acid sequences contained arginine‒glycine or arginine‒glycine‒glycine motifs and eight conserved motifs belonging to the DEAD‒box protein family, and showed highly similarity and identity with vasa homologs of other teleosts. Tissue specific expressions of Tp‒ and Gp‒vasa were detected in testis and ovary. In situ hybridization analysis showed that Tp‒ and Gp‒vasa mRNA expressed in oogonia and oocytes in the ovary, and spermatogonia in the testis, while it was not detected in the primary and secondary spermatocytes, spermatid, spermatozoa or gonadal somatic cells. These findings suggest that Tp‒ and Gp‒vasa should be useful for studies of germ cells specification and differentiation in Takifugu puffers.