Abstract

Class III chitin synthases in filamentous fungi are important for hyphal growth and differentiation of several filamentous fungi. A genomic clone containing the full gene encoding Chs4, a class III chitin synthase in Penicillium chrysogenum, was cloned by PCR screening and colony hybridization from the genomic library. Nucleotide sequence analysis and transcript mapping of chs4 revealed an open reading frame (ORF) that consisted of 5 exons and 4 introns and encoded a putative protein of 915 amino acids. Nucleotide sequence analysis of the 5?lanking region of the ORF revealed a potential TATA box and several binding sites for transcription activators. The putative transcription initiation site at ?6 position was identified by primer extension and the expression of the chs4 during the vegetative growth was confirmed by Northern blot analysis. Amino acid sequence analysis of the Chs4 revealed at least 5 transmembrane helices and several sites for post-translational modifications. Comparison of the amino acid sequence of Chs4 with those of other fungi showed a close relationship between P. chr ysogenum and genus Aspergillus.

• Keywords: Penicillium chrysogenum; chitin synthase gene; screening; genomic structure; transcript mapping