A full-length cDNA, MIC-3, has been identified from a lambda ZAPII cDNA library constructed from the mRNA of nematode-resistant cotton (Gossypium hirsutum L.) roots after infection with root-knot nematode (Meloidogyne incognita). The putative open reading frame of MIC-3 encoded a protein of 141 amino acids with a calculated molecular mass of 15.3 kDa. Seven alternative polyadenylation sites have been identified for the MIC-3 transcripts, and the major transcripts are the longest ones. The MIC-3 gene contains a single intron within its coding region and belongs to a novel, multi-gene family containing up to six members. Expression of MIC-3 is root localized and specifically enhanced in the nematode induced, immature galls of resistant cotton line M-249, suggesting that MIC-3 may play a critical role in the resistance response to root-knot nematode.