Sequencing of a large number of expressed sequence tags from Blastocladiella emersonii revealed the presence of ten distinct putative members of the 70 kDa-heat shock protein (Hsp70) family in this fungus. The amino acid sequence deduced from eight of these cDNAs showed significant similarity to members of the Saccharomyces cerevisiae Hsp70 family, and the remaining displayed high sequence homology with hsp70 gene products from other organisms. The hsp70-3 gene was the most highly expressed at normal temperatures and was poorly induced during heat shock. Except for hsp70-4 and hsp70-6, all other hsp70 genes were induced to different degrees upon exposure of B. emersonii cells to heat shock, with hsp70-1 gene presenting the highest transcript levels. Phylogenetic analysis of complete B. emersonii putative Hsp70 protein sequences indicated that Hsp70-1 and Hsp70-3 corresponded to cytosolic proteins, whereas Hsp70-7 and Hsp70-9 are probably localized in the endoplasmic reticulum and mitochondria, respectively.