ISSN:
1617-4623
Keywords:
Key words Penicillium italicum
;
Lanosterol 14α-demethylase
;
Fungicides
;
Sterol biosynthesis inhibitors
;
Cytochrome P450
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract The CYP51 gene encoding eburicol 14α-demethylase (P45014DM) was cloned from a genomic library of the filamentous fungal plant pathogen Penicillium italicum, by heterologous hybridisation with the corresponding gene encoding lanosterol 14α-demethylase from the yeast Candida tropicalis. The nucleotide sequence of a 1739-bp genomic fragment and the corresponding cDNA clone comprises an open reading frame (ORF) of 1545 bp, encoding a protein of 515 amino acids with a predicted molecular mass of 57.3 kDa. The ORF is interrupted by three introns of 60, 72 and 62 bp. The C-terminal part of the protein includes a characteristic haem-binding domain, HR2, common to all P450 genes. The deduced P. italicum P45014DM protein and the P45014DM proteins from Candida albicans, C. tropicalis and Saccharomyces cerevisiae share 47.2, 47.0 and 45.8% amino acid sequence identity. Therefore, the cloned gene is classified as a member of the CYP51 family. Multiple copies of a genomic DNA fragment of P. italicum containing the cloned P450 gene were introduced into Aspergillus niger by transformation. Transformants were significantly less sensitive to fungicides which inhibit P45014DM activity, indicating that the cloned gene encodes a functional eburicol 14α-demethylase.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF02172984
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