PCR was used to amplify fragments corresponding to the chitin synthase () genes from the Oomycetes Saprolegnia monoica, and utilizing as primers, oligonucleotides designed from the conserved region of genes of chitinous fungi. Chitin synthase homologues were found in the three cellulosic fungi. The chitin synthase 2 gene () from was cloned, sequenced and characterized. The amino acid sequence deduced from the genomic DNA revealed several domains, corresponding to the catalytic domains and polypeptide signatures, of high identity with genes from chitinous fungi. Existence of a gene family in was supported by the identification of two sequences among the PCR products, the localization of homologues on two chromosomes, and the detection of two transcripts in mycelia and protoplasts. Polyclonal anti-chitin synthase antibodies raised against the N-terminal and the neutral fragments of the products revealed, respectively, two and four proteins in membrane fractions and a truncated active form in entrapped product. The overall comparison of the structure and organization of genes indicates that in spite of their divergent evolution, Oomycetes and chitinous fungi have evolved with conserved chitin synthase systems.


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