The T-box genes, including Brachyury, encode a novel family of transcription factors that play critical roles in various processes of development, in particular, mesoderm formation in chordate embryos. In the case of the ascidian Halocynthia roretzi, the Brachyury (As-T) is expressed exclusively in notochord cells, whereas another T-box gene (As-T2) is expressed in the muscle cells and in the tip of the tail of tailbud embryos. In a previous study, we suggested that the combined pattern of the spatial expressions of As-T and As-T2 appears to correspond to that of a single vertebrate Brachyury gene (Yasuo et al., Dev Biol 1996;180:773-779). The present molecular phylogenetic analysis suggested that As-T2 is a divergent member of the T-box family with a similarity to the Tbx6 subfamily. Vertebrate members of this subfamily are expressed in the posterior paraxial mesoderm, and the mouse Tbx6 is essential for the specification of posterior somite. To investigate the function of As-T2, we examined an ectopic- and/or overexpression of this gene by injecting synthetic mRNA into fertilized eggs. The results showed that the injection of As-T2 mRNA induced an ectopic expression of muscle-specific myosin heavy-chain gene and actin gene, especially in presumptive epidermal cells. This ectopic muscle-specific expression was accompanied by the partial suppression of an epidermis-specific gene expression. The overexpression of As-T2, however, rarely affected the expression of As-T (Brachyury) and genes that are expressed in the tailbud.