Isolation of a cDNA encoding a 31-kDa, pathogenesis-related 5/thaumatin-like (PR5/TL) protein abundantly expressed in apple fruit (Malus domestica cv. Fuji)

Abstract

A fruit-specific and pathogenesis-related 5/thaumatin-like (PR5/TL), 31-kDa protein was isolated by 2D-PAGE from fully-grown apples (Malus domestica cv. Fuji) and named Mdt11 (Malus domestica thaumatin-like protein 1). Using the N-terminal sequence of the protein, the full-length cDNA encoding Mdt11 was isolated. The cDNA clone (Mdt11) consists of 944 bp with an open reading frame (ORF) of 744 bp encoding a protein of 247 amino acids. The deduced amino acid sequence of Mdt11 shows high similarity to the sequences of PR5/TL proteins. Mdt11 is a slightly acidic protein with a putative signal peptide and a putative N-glycosylation site, and lacks a C-terminal extension. This suggests that Mdt11 is an apoplastic glycoprotein. Results of northern blotting indicated that expressions of Mdt11 are developmentally regulated. Southern blot analysis showed that Mdt11 may be present as a single copy, and there exist other genes closely related to Mdt11 in the apple genome.