Targeted gene replacement by homologous recombination in Drosophila stimulates production of second-site mutations.
Gene replacement by homologous recombination is a powerful technique for generating mutations in Drosophila. while using this technique for the BEAF gene, we encountered non-targeted lethal mutations on the target chromosome that complicated the analysis of the BEAF mutations until they were discovered and removed by meiotic recombination. Subsequent experiments indicated that the gene-targeting method leads to a modest but significant three-fold increase in the rate of production of non-targeted lethal mutations. It is important to be aware of this phenomenon when using this method.
Roy
Swarnava
S
NIDDK, Metabolic Diseases Branch, National Institutes of Health, Bethesda, MD, USA. roys2@niddk.nih.gov
Hart
Craig M
CM
D000818:Animals; D004268:DNA-Binding Proteins / Q000235:genetics; D004330:Drosophila / Q000235:genetics*; D029721:Drosophila Proteins / Q000235:genetics; D005136:Eye Proteins / Q000235:genetics; D018390:Gene Targeting; D016296:Mutagenesis; D011995:Recombination, Genetic
C452919:BEAF-32 protein, Drosophila; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D005136:Eye Proteins