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Prothymosin alpha (ProTα) gene of Giant Pandas, which encodes Prothymosin alpha of Giant Pandas(P-ProTα), was cloned from Giant Pandas' spleen for the first time,. Bioinformatics analysis was performed to predict the characteristics of P-ProTα. The results indicated that Prothymosin alpha gene encodes a polypeptide, molecular mass of 12.0709Kda and comprising 110 amino acids, without signal peptide and the transmembrane region. The phylogenetic tree showed that P-ProTα was similar to some mammalian species, but clustered within a monophyletic clade. By multiple sequence alignment, what we found was that ProTα was as the conserved sequence existing in mammalian species. The online analysis of the physico-chemical properties indicated that the protein possessed typical characteristics of ProTα lacking aromatic and sulfur amino acids and being highly acidic. These results will provide a fundament for further biological functional study of the Prothymosin alpha of Giant Pandas and physiological features of the ProTα gene of Giant Pandas.