By cloning and sequencing cDNA, the primary structure of a mycelial aggregate-specific lectin of Pleurotus cornucopiae was determined. The amino acid sequence was novel and elucidated unique properties of this lectin: It was composed of 373 amino acids. 33 of which constitute a signal sequence. The sequence of the mature lectin consisted of two homologous regions having five glycosylation recognition signals and six cysteine residues. However, the distribution of these elements in the two regions was biased. Expression of cDNA in Escherichia coli and Pichia pastoris revealed the requirement of glycosylation to produce the functional lectin. Gel filtration followed by gel electrophoretic analyses of the purified lectin showed that the active component moved faster than the bulk of the protein, suggesting that the most active lectin formed an oligomer of subunits through disulfide bonds. From these observations, a model for the structure of the active form of this lectin is proposed. Southern hybridization using the cDNA as a probe revealed the presence of several genes. The lectin gene was composed of five exons and five introns.