title: Fluorescence Assisted Selection of Transformants (FAST): Using flow cytometry to select fungal transformants.
abstract: The availability of drug resistance markers for fungal transformation is often a limiting factor in both fungal genetics research and industrial applications. We describe a new technique using flow cytometry to select fungal transformants using well-known fluorescent proteins as markers for transformation. This new technique, Fluorescence-Assisted Selection of Transformants (FAST), was used for a transformation of Fusarium oxysporum with GFP as a marker targeted at a specific site on chromosome 14. The resulting strain was then transformed again with a gene replacement construct containing both RFP and a gene for drug resistance as markers. By directly comparing FAST with drug resistance selection we show that both methods yield comparable numbers of gene deletion mutants.
authors: Molecular Plant Pathology, University of Amsterdam, Amsterdam, The Netherlands.
authors: Ido
authors: I
authors: Vlaardingerbroek
authors: Molecular Plant Pathology, University of Amsterdam, Amsterdam, The Netherlands.
authors: Bas
authors: B
authors: Beerens
authors: Molecular Plant Pathology, University of Amsterdam, Amsterdam, The Netherlands.
authors: Shermineh
authors: S
authors: Shahi
authors: Molecular Plant Pathology, University of Amsterdam, Amsterdam, The Netherlands. Electronic address: m.rep@uva.nl.
authors: Martijn
authors: M
authors: Rep
mesh_terms: D005434:Flow Cytometry / Q000379:methods*; D005453:Fluorescence; D005656:Fungal Proteins / Q000737:chemistry*; D005670:Fusarium / Q000235:genetics*; D014170:Transformation, Genetic
chemical_list: D005656:Fungal Proteins
keywords: Flow cytometry; Fluorescent proteins; Fusarium; Transformation