Cloning and expression of squalene epoxidase from the pathogenic yeast Candida albicans
Détails
ID Serval
serval:BIB_0504EEBA594F
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Cloning and expression of squalene epoxidase from the pathogenic yeast Candida albicans
Périodique
Gene
ISSN
0378-1119 (Print)
Statut éditorial
Publié
Date de publication
04/1997
Volume
189
Numéro
1
Pages
119-26
Notes
Comparative Study
Journal Article --- Old month value: Apr 11
Journal Article --- Old month value: Apr 11
Résumé
The allylamine antimycotic terbinafine prevents the formation of sterols by specifically inhibiting squalene epoxidase (SE). The biological and biochemical action of terbinafine on fungal pathogens has been well investigated, but little is known at the molecular level. Here we report the cloning, sequencing and expression of the target of terbinafine from the major pathogen Candida albicans. A C. albicans genomic DNA library was constructed in gamma ZAP Express and screened with a DNA fragment obtained by polymerase chain reaction with two primers designed from sequences common to Saccharomyces cerevisiae and rodent SEs. Two types of clone, approximately 3.9 kbp and 4.1 kbp, were isolated. Both contained an identical open reading frame of 1488 nucleotides, while a few sequence differences were found in the flanking regions, suggesting an allelic heterogeneity. The deduced protein sequence of C. albicans SE, 496 amino acids (55324 Da), is 54% and 41% identical to those of S. cerevisiae and rat, respectively. A 1.8-kb transcript was observed on Northern blots of C. albicans mRNA. Polyclonal antibodies, raised against an internal peptide of C. albicans SE, recognized a protein associated with the particulate fraction of M(r) 55000 on Western blots of C. albicans extracts. C. albicans SE was overexpressed in S. cerevisiae with the expression vector pYES2. In homogenates from S. cerevisiae overexpressing the C. albicans protein SE activity was 10-fold higher than the endogenous activity from controls.
Mots-clé
Amino Acid Sequence
Animals
Base Sequence
Candida albicans/*enzymology/*genetics/pathogenicity
Cloning, Molecular
Fungal Proteins/*genetics/isolation & purification
Gene Expression Regulation, Fungal
Genetic Vectors/chemistry
Immunoblotting
Mice
Molecular Sequence Data
Oxygenases/*genetics/isolation & purification
Rats
Saccharomyces cerevisiae/genetics
Sequence Alignment
Squalene Monooxygenase
Transcription, Genetic
Pubmed
Web of science
Création de la notice
25/01/2008 17:32
Dernière modification de la notice
20/08/2019 13:26