Abstract: AIM A cDNA expressional library from the caudal spine of Dasyatis akajei was constructed into pcDNA 3.0 eukaryotical expression plasmid. METHODS SMART^TM protocol was used for cDNA Library Construction and bioinformatics analysis was carried out. RESULTS Fifty six novel EST clones were obtained from the library, in which there were 24 full-length clones, including IPL tumor suppressor gene, cyclophilin A, selenoprotein W, cystatin A, tryosyl-tRNA synthetase and calcineurin homologous protein. CONCLUSION These genes were mostly first reported in Dasyatis akajei. Further studies may partly explain the molecular mechanisms by which the caudal spine of Dasyatis akajei were often used in the treatment of inflammation, parasitosis and cancer in the traditional Chinese medicine.