Abstract: By RACE-PCR cloning, a cDNA encoding taxane 2α-O-benzoyltransferase (designated as TmTBT) was isolated from Taxus media, which catalyzes the conversion of 2-debenzoyl-7,13-diacetylbaccatin III, a semisynthetic substrate, to 7,13-diacetylbaccatin III. The full-length cDNA of TmTBT was 1478 bp and contained a 1320 bp Open Reading Frame (ORF) encoding a protein of 440 amino acid residues with molecular weight of 50,090 Da and an isoelectric point (pI) of 6.25, similar to taxane 2α-O-benzoyltransferase from Taxus cuspidata. Sequence comparison analysis revealed that TmTBT had high similarity with other members of plant transferase family. Phylogenetic tree analysis showed that TmTBT had close relationship with taxane 2α-O-benzoyltransferase from T. cuspidata. Tissue expression pattern analysis revealed that TmTBT expressed only in leaves and no expression could be detected in fruits and stems, indicating that TmTBT was a tissue-specific gene.