To elucidate the diversity of troponin C (TnC) during chordate evolution, we determined the organization of TnCs from the amphioxus, the lamprey, and the frog. Like the ascidian, the amphioxus possesses a single gene of TnC, and the fundamental gene structure is identical with the ascidian TnC. However, because alternative splicing does not occur in amphioxus, the potential for generation of TnC isoforms through this event arises only in the ascidian lineage. From the frog Xenopus laevis, two distinct cDNAs encoding fTnC isoforms and a single s/cTnC cDNA were determined. The duplication of the fTnC gene may be a character of only Xenopus or closely related species. The lamprey possesses two cDNAs each encoding fTnC and s/cTnC. The lamprey is the earliest diverged species among vertebrates, and thus it is supposed that the presence of both fTnC and s/cTnC is universal among vertebrate species, and that the gene duplication might have occurred at a vertebrate ancestor after the protochordate/vertebrate divergence. The position of the 4th intron is 3.24/0 in protochordate TnC genes, but at 3.11/2 in vertebrate fTnCs and s/cTnCs. It is suggested that the 4th intron sliding might have occurred prior to the gene duplicatio
