Abstract: Cadmium (Cd) of tobacco is a pollutant that is extremely toxic to the health of humans. Cadmium/zinc-transporting ATPase HMA1 gene has been characterized to function in the plant cadmium/zinc-transporting. The complete coding sequence of tobacco cadmium/zinc-transporting ATPase HMA1 gene was amplified by RT-PCR. The open reading frame of tobacco cadmium/zinc-transporting ATPase HMA1 gene was 2418 bp which encodes a protein of 805 amino acids. BLAST analysis revealed that tobacco cadmium/zinc-transporting ATPase HMA1 protein shares high homology with the cadmium/zinc-transporting ATPase HMA1 of potato (85%), Lycopersicon esculentum (84%), wine grape (71%), sweet orange (71%), soybean (69%) and thale cress (68%). Results also showed that tobacco cadmium/zinc-transporting ATPase HMA1 gene has a closer genetic relationship with the cadmium/zinc-transporting ATPase HMA1 gene of Lycopersicon esculentum. Prediction of transmembrane helices showed that tobacco cadmium/zinc-transporting ATPase HMA1 might be a transmembrane protein. The expression profile was studied and results indicated that tobacco cadmium/zinc-transporting ATPase HMA1 gene was moderately expressed in root, leaf and stem but hardly expressed in flower. These results established the primary foundation of utilizing tobacco cadmium/zinc-transporting ATPase HMA1 gene to descrease the cadmium content of tobacco and benefit the health of humans in the future.