Identification, purification and gene cloning of a protein from Tuber dryophilum fruitbodies homologous to TBF-1 protein

Abstract

This paper reports a study aimed at gaining new information on the molecular composition of fruitbodies of Tuber spp. The presence of proteins homologous to TBF-1, which is highly specific for the fruitbody-phase of Tuber borchii, has been investigated in other white truffles. SDS-PAGE analyses revealed that only Tuber dryophilum fruitbodies possess a similar protein. This protein was purified by HPLC and partially sequenced, confirming a high degree of homology with TBF-1. Several PCR analyses of the genomic DNA, were performed to evaluate whether the absence of proteins homologous to TBF-1 in other white truffle species was a result of the absence of the coding genes. T. dryophilum gave an amplification product corresponding to the entire gene (tdf-1), but no products were obtained from the other species. Tdf-1 was sequenced and its organisation studied since it is one of the first genes isolated from a Tuber species. The deduced amino acid sequence was compared to that of TBF-1, to evaluate the presence of conserved regions, in an attempt to gain new information about their role in fruitbody formation.