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Cloning and Sequence Analysis of the Eburicol 14 α ‐Demethylase Encoding Gene ( CYP51 ) from the Japanese Pear Scab Fungus Venturia nashicola
Author(s) -
Cools H. J.,
Ishii H.,
Butters J. A.,
Hollomon D. W.
Publication year - 2002
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1046/j.1439-0434.2002.00776.x
Subject(s) - biology , genomic dna , demethylase , gene , polymerase chain reaction , cloning (programming) , genetics , demethylation , molecular cloning , pear , dna , sequence analysis , microbiology and biotechnology , botany , complementary dna , dna methylation , gene expression , epigenetics , computer science , programming language
The slow growth of many filamentous fungi in vitro prevents rapid isolation of large quantities of genomic DNA, thereby hampering molecular studies. Using a polymerase chain reaction (PCR)‐based adapter ligation cloning technique that requires only nanogram quantities of genomic DNA we have successfully cloned the complete CYP51 gene encoding the eburicol 14 α ‐demethylase, the target for DMI (Demethylation‐inhibiting) fungicides, from the Japanese pear scab fungus, Venturia nashicola . Analysis of predictedamino acid sequences of CYP51s from strains less sensitive to DMIs revealed no alterations when compared to the sensitive reference strain.