Abstract: Sucrose phosphate synthase (SPS) gene from cassava RS01 was cloned to study its biological functions. The conserved fragment of the gene was first obtained by homologous cloning, and its full length cDNA by rapid amplification of cDNA ends (RACE). The characteristics, gene sequence and other biological information retrieved were analyzed. Expressions of the genes from various tissues of the plants at different times were compared by RT-qPCR. The results showed that the cDNA of SPS gene had a full length of 3, 857 bp, and an ORF of 3, 228 bp encoding a protein of 1, 076 amino acids. It was subsequently named MeSPS with a GenBank number of KX822780. Its homologous alignments with the nucleotide and protein databases were highly homologs with those of Jatropha carcas L., Ricinus communis L., and Litchi chinensis Sonn. at 89%, 89%, and 87%, respectively. Its amino acid sequence was 77%-92% in similarity with the SPS genes of other plants. The qRT-PCR analysis showed that MeSPS gene was highly expressed during seeding, and, was more expressed in the leaves than the stems or roots on a plant in a same growth period.