Pathogenesis-related (PR) proteins, induced in plants in response to various biotic and abiotic stresses, have been assumed to play a role in plant defense system. Proteins of the PR5 family, also named thaumatin-like proteins (TLPs), have been detected in numerous plant species. In this research, a novel PR5 gene, designated as TaPR5, was isolated and characterized from wheat leaves (cv. Suwon 11) infected by the stripe rust pathotype CY23 (incompatible interaction) using the rapid amplification of cDNA ends (RACE). TaPR5 was predicted to encode a protein of 173 amino acids with an estimated molecular mass of 17.6 kDa and a theoretical pI of 4.64. The deduced amino acid sequence of TaPR5 showed a significant sequence similarity with PR5 and TLPs from barley and other plants and contained a putative signal peptide at the amino terminus. Southern blot analysis indicated that TaPR5 is coded by a single-copy gene. Quantitative real-time polymerase chain reaction (qRT-PCR) analyses revealed that TaPR5 transcript is significantly induced and upregulated in the incompatible interaction while in the compatible interaction a relative low level of the transcript was detected. TaPR5 was also induced by phytohormones (SA, JA and ABA) and stress stimuli (wounding, cold temperature and high salinity). Using an assay of onion epidermal cells indicated accumulation of TaPR5 protein in the apoplast. The immunocytochemical method showed that the TaPR5 protein was detected on cell walls of wheat leaves in the incompatible interaction at markedly higher labeling density compared with the compatible interaction.