Detection of gene-disruption events in Aspergillus transformants by polymerase chain reaction direct from conidiospores.
We describe a rapid method for the identification of gene disruption events after DNA-mediated transformation of Aspergillus fumigatus. This involves a polymerase chain reaction in which the target DNA is added in the form of intact conidiospores. Using one primer specific to the plasmid DNA and a second primer specific to the target gene on the chromosome, it is possible to identify gene disruption events among the more common ectopic integrations approximately 4 h after sporulating transformants appear on selective medium.
Aufauvre-Brown
A
A
Department of Infectious Diseases and Bacteriology, Royal Postgraduate Medical School, Hammersmith Hospital, London, UK.
Tang
C M
CM
Holden
D W
DW
D001232:Aspergillus fumigatus / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D004271:DNA, Fungal; D008969:Molecular Sequence Data; D016296:Mutagenesis; D016133:Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000235:genetics; D013172:Spores, Fungal; D014170:Transformation, Genetic
D004271:DNA, Fungal; D012697:Serine Endopeptidases; C026109:oryzin