Aux/IAA genes are a large gene family in plant, many of which are rapidly and specifically induced by auxin. Previous data have illustrated that Aux/IAA genes participated in both auxin signaling and plant development. In order to discover the biofunction of SlIAA3 gene, an Aux/IAA gene from tomato, we isolated the full-length cDNA and the corresponding genomic DNA of this gene. Sequence analysis results showed that there were two introns and three extrons in SlIAA3 gene. DNA gel-blot analysis revealed that SlIAA3 was a single copy in tomato and SlIAA3 was bin-mapped in chromosome 9-G region using 75 tomato introgression lines. Expression analysis showed that SlIAA3 was expressed in all tissues tested, whereas the levels of transcript abundance were different. The expression patterns indicating that SlIAA3 gene should be involved in the root development and auxin signaling.