Print Email Facebook Twitter Isolation and molecular characterisation of the gene encoding eburicol 14α-demethylase (CYP51) from Penicillium italicum Title Isolation and molecular characterisation of the gene encoding eburicol 14α-demethylase (CYP51) from Penicillium italicum Author van Nistelrooy, J.G.M. van den Brink, J.M. van Kan, J.A.L. van Gorcom, R.F.M. de Waard, M.A. Prins Maurits Laboratorium TNO Publication year 1996 Abstract The CYP51 gene encoding eburicol 14α-demethylase (P450(14DM)) 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 (ORE) 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 P450(14DM) protein and the P450(14DM) 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 P450(14DM) activity, indicating that the cloned gene encodes a functional eburicol 14α-demethylase. Subject Biology14α-demethylaseCytochrome P450FungicidesPenicillium italicumSterol biosynthesis inhibitorsCytochrome p450 isoenzymeEnilconazoleEtaconazoleFenarimolFungicideUnclassified drugArticleControlled studyGene isolationGenetic analysisMolecular cloningMolecular geneticsNonhumanPenicilliumPriority journalSterol synthesisYeastAmino Acid SequenceBase SequenceBlotting, NorthernCloning, MolecularCytochrome P-450 Enzyme SystemDNA PrimersFungal ProteinsFungicides, IndustrialGenes, FungalLanosterolMolecular Sequence DataOxidoreductasesPenicilliumPyrimidinesRestriction MappingSequence Analysis, DNASequence Homology, Amino Acid To reference this document use: http://resolver.tudelft.nl/uuid:510be056-261e-41ca-bf1c-de636f736526 DOI https://doi.org/10.1007/s004380050126 TNO identifier 233310 ISSN 0026-8925 Source Molecular and General Genetics, 250 (6), 725-733 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.