Abstract: To clone glutathione S-transferase gene（GST）of Saccharum spontaneum L. In this study，a GST gene from S. spontaneum L. was cloned by in silico cloning and RT-PCR，named as SsGST，characteristics of SsGST gene were analyzed using bioinformatics tools. Sequence analysis showed that the full length cDNA of SsGST was 702 bp，which encoded 224 amino acids. The predicted protein molecular formula was C1119H1756N300O327S5，whose molecular mass was 24.8 ku and isoelectric point was 6.21. Hydrophobic analysis showed that SsGST protein was a hydrophilic protein. Conserved domain prediction indicated that SsGST protein had GST-N domain and GST-C domain. SsGST protein sequence had higher similarities with the GST protein sequence of maize，sorghum，foxtail millet and sugarcane and so on. Phylogenetic analysis showed that SsGST protein was close to GST protein of maize.