title: A novel, negative selectable marker for gene disruption in Dictyostelium.
abstract: The expression of an ochre suppressor mutant of the GluII(UUA) tRNA appears to be lethal to Dictyostelium, and offers a novel 'positive negative' strategy to select for targeted gene disruption by homologous recombination. Inclusion of the suppressor tRNA gene decreases the overall transformation frequency by approximately 20-fold. This increases the proportion of targeted gene disruptions to over 90%.
authors.affiliation: The Imperial Cancer Research Fund, Clare Hall Laboratory, South Mimms, Herts, UK.
authors.forename: A
authors.initials: A
authors.lastname: Morrison
authors.forename: R
authors.initials: R
authors.lastname: Marschalek
authors.forename: T
authors.initials: T
authors.lastname: Dingermann
authors.forename: A J
authors.initials: AJ
authors.lastname: Harwood
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D004023:Dictyostelium / Q000235:genetics*; D018390:Gene Targeting / Q000379:methods*; D005804:Genes, Lethal; D016153:Genes, Suppressor; D005819:Genetic Markers; D012352:RNA, Transfer, Glu / Q000235:genetics*; D011995:Recombination, Genetic; D014170:Transformation, Genetic
chemical_list: D005819:Genetic Markers; D012352:RNA, Transfer, Glu