PCR Cloning and Detection of Point Mutations in the Eburicol 14alpha-demethylase (CYP51) Gene From Erysiphe Graminis F. Sp. Hordei, a "Recalcitrant" Fungus

Curr Genet. 1998 Dec;34(5):399-403. doi: 10.1007/s002940050413.

Abstract

Molecular studies of some micro-organisms are hampered by the difficulty of obtaining sufficient amounts of nucleic acids. A cloning strategy based on PCR has therefore been used to clone the eburicol 14alpha-demethylase (CYP51) gene of the obligate fungus Erysiphe graminis f. sp. hordei (Egh) using minute amounts of genomic DNA. The CYP51 gene encodes the enzymatic target of a major group of fungicides. Sequencing CYP51 from different Egh isolates revealed the occurrence of two alleles for this gene. An allele-specific PCR assay was developed to detect each CYP51 allele.

MeSH terms

  • Alleles
  • Amino Acid Sequence
  • Ascomycota / chemistry
  • Ascomycota / enzymology
  • Ascomycota / genetics*
  • Cloning, Molecular
  • Cytochrome P-450 Enzyme System / genetics*
  • DNA, Complementary / chemistry
  • DNA, Complementary / genetics
  • DNA, Fungal / chemistry
  • DNA, Fungal / genetics
  • Gene Amplification
  • Genes, Fungal / genetics*
  • Genetic Variation
  • Molecular Sequence Data
  • Oxidoreductases / genetics*
  • Point Mutation
  • Polymerase Chain Reaction
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Sterol 14-Demethylase

Substances

  • DNA, Complementary
  • DNA, Fungal
  • Cytochrome P-450 Enzyme System
  • Oxidoreductases
  • Sterol 14-Demethylase

Associated data

  • GENBANK/AF052515