Gene deletion and allelic replacement in the filamentous fungus Podospora anserina.
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 DeltaPaKu70 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.
El-Khoury
Riyad
R
CNRS, Centre de Génétique Moléculaire, UPR 2167, Avenue de la Terrasse, Bat. 26, 91198, Gif-sur-Yvette, France.
Sellem
Carole H
CH
Coppin
Evelyne
E
Boivin
Antoine
A
Maas
Marc F P M
MF
Debuchy
Robert
R
Sainsard-Chanet
Annie
A
D000483:Alleles; D034961:Antigens, Nuclear / Q000235:genetics; D015139:Blotting, Southern; D004268:DNA-Binding Proteins / Q000172:deficiency / Q000235:genetics; D017353:Gene Deletion; D018390:Gene Targeting; D005800:Genes, Fungal / Q000235:genetics*; D000072200:Ku Autoantigen; D048468:Podospora / Q000235:genetics*
D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D000072200:Ku Autoantigen