title: Heat shock at an elevated temperature improves transformation efficiency of protoplasts from Podospora anserina.
abstract: We have developed an improved transformation procedure for the filamentous fungus Podospora anserina. This procedure is based on the observation that a heat shock at an elevated temperature (48 degrees C) improves the competence of P. anserina protoplasts for transformation 5- to 10-fold. This is observable only if the heat shock is applied before the addition of transforming DNA. An increase in competence is observed immediately after the heat shock, and heat-shocked cells are still competent after 20-30 min. The mechanism by which heat shock improves competence remains unclear. The modified transformation procedure gives as many as 200-500 stable transformants per microgram of plasmid DNA containing the P. anserina ura5 gene. This should allow direct cloning of P. anserina genes from a cosmid library.
authors.affiliation: Laboratoire de Génétique, Université de Bordeaux II, Talence, France.
authors.forename: T
authors.initials: T
authors.lastname: Bergès
authors.forename: C
authors.initials: C
authors.lastname: Barreau
mesh_terms: D001203:Ascomycota / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics; D005800:Genes, Fungal; D006358:Hot Temperature; D011523:Protoplasts; D014170:Transformation, Genetic; D013098:Xylariales / Q000235:genetics*
chemical_list: D004271:DNA, Fungal