title: The Cre/lox system: a practical tool to efficiently eliminate selectable markers in fungal endophytes.
abstract: Interspecific hybridization is a common evolutionary process for the many epichloid fungi that consequently possess multiple gene copies accumulated from their parental strains. Serial gene manipulations in such strains are impeded by the limited availability of selectable resistance marker genes. Therefore, we developed a method for marker elimination suitable for a range of filamentous fungi that allows the reuse of the same marker for successive manipulations, and can also generate gene knockout mutants free of any foreign genes. For epichloae, the complete elimination of the marker gene from the genome would mitigate public concerns and regulatory hurdles to the use of such fungal strains in field experiments.
authors.affiliation: Department of Plant Pathology, UK-Advanced Genetic Technologies Center, University of Kentucky, Lexington, KY, USA.
authors.forename: Simona
authors.initials: S
authors.lastname: Florea
authors.forename: Caroline
authors.initials: C
authors.lastname: Machado
authors.forename: Kalina
authors.initials: K
authors.lastname: Andreeva
authors.forename: Christopher L
authors.initials: CL
authors.lastname: Schardl
mesh_terms: D017931:DNA Primers / Q000235:genetics; D004270:DNA, Circular / Q000235:genetics; D060026:Endophytes / Q000235:genetics* / Q000378:metabolism; D018014:Gene Transfer Techniques; D005819:Genetic Markers / Q000235:genetics; D019426:Integrases / Q000378:metabolism*; D055328:Neotyphodium / Q000235:genetics* / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011523:Protoplasts / Q000378:metabolism; D014170:Transformation, Genetic / Q000235:genetics*
chemical_list: D017931:DNA Primers; D004270:DNA, Circular; D005819:Genetic Markers; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase; C045073:Cre recombinase; D019426:Integrases