A novel, negative selectable marker for gene disruption in Dictyostelium.
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%.
Morrison
A
A
The Imperial Cancer Research Fund, Clare Hall Laboratory, South Mimms, Herts, UK.
Marschalek
R
R
Dingermann
T
T
Harwood
A J
AJ
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
D005819:Genetic Markers; D012352:RNA, Transfer, Glu