Trehalose, an important disaccharide in fungal spores, plays important role in protects against many environmental stresses, such as heat and salt. In this study, a gene (PiTPS) encoded trehalose-6-phosphate synthase was isolated from Phellinus igniarius (L.: Fr.) Quél, a basidiomycete fungus belonging to the family Polyporaceae. It contains many bioactive compounds that have been reported to possess antiviral, antibacterial, antitumor and antimutagenic activities. The PiTPS cDNA is composed of 2688 nucleotides, which encodes a protein of 517 amino acids and its molecular mass is about 100 kDa. The amino acid sequence has a relatively high homology with the TPS in several other filamentous fungi. The predicted protein contains a glycol_transf_20 domain and a trehalose_PPase domain. Genomic structure of PiTPS is composed of three exons with 607, 763 and 1,318bp and two introns with 73 and 52 bp. In this rearch, we clone and express the TPS gene in E. coli, constructed engineering strain, and laid the foundation for further study its structure and function.