Journal
CURRENT GENETICS
Volume 53, Issue 4, Pages 249-258Publisher
SPRINGER
DOI: 10.1007/s00294-008-0180-3
Keywords
filamentous fungi; Podospora anserina; homologous recombination; Ku70; knock-out mutant; allelic replacement
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Gene replacement via homologous recombination is a fundamental tool for the analysis of gene function. However, this event is rare in organisms like the filamentous fungus Podospora anserina. We show here that deletion of the PaKu70 gene is an efficient strategy for improving gene manipulation in this organism. By using the Delta PaKu70 strain, it is now possible (1) to produce deletion mutants with an efficiency of 100%, (2) to achieve allelic exchange by introducing a mutated allele associated with a selection cassette at the locus, (3) to introduce a mutation in a gene without co-insertion of a selectable marker and without any modification of the target locus.
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