The adenine nucleotide translocase (ANT) is the most abundant mitochondrial inner membrane protein, which catalyzes the exchange of ADP and ATP between cytosol and mitochondria and participates in many models of mitochondrial apoptosis. In the present study, the full sequence of P. monodon ANT gene was cloned and named PmANT. The full length cDNA of PmANT contained a 5’ untranslated region (UTR) of 65bp, a 3’ UTR of 393bp and an ORF of 930bp encoding a polypeptide of 309 amino acids with an estimated molecular mass of 33.622kD. As other animal ANTs, the structure of the PmANT protein consists of three homologous repeated domains. But there are not signal peptide and glycosylation sites in PmANT protein. Sequence alignment analysis showed that the PmANT with the ANT of Litopenaeus vannamei shared a similarity of 98.7% and the homology of 97.4%. Analysis of the tissue expression pattern of the PmANT showed that the PmANT mRNA was expressed in all tested tissues, including heart, ovary, testis, muscle, eye disease nerve, cranial nerve, lymphoid tissue, stomach, intestines, hepatopancreas, gill, hemocyte, with the highest levels in muscle of male and the lowest level in immature testis. Furthermore, the PmANT expression was found to be a higher level in mature testis than in immature testis. The PmANT expression was found in the six ovarian stages of development and the expression of ovarian growth levels in the third growth phase was significantly higher than that of others except the sixth growth phase, while lowest in the fourth growth phase. The study provided essential materials and laid a firm foundation for the further research of the function of PmANT in gonad development of P. monodon.