Facilitated isolation of rare recombinants by ligase chain reaction: selection for intragenic crossover events in the Drosophila optomotor-blind gene

• Published in: Molecular & general genetics Vol. 245; no. 6; p. 734
• Main Authors: Balles, J, Pflugfelder, G O
• Format: Journal Article
• Language: English
• Published: Germany 15.12.1994
• Subjects: Animals; Base Sequence; DNA Ligases - metabolism; DNA-Binding Proteins - genetics; Drosophila melanogaster - genetics; Drosophila Proteins; Genes, Insect; Molecular Sequence Data; Nerve Tissue Proteins - genetics; Nucleic Acid Hybridization; Point Mutation; Recombination, Genetic; T-Box Domain Proteins
• ISSN: 0026-8925
• DOI: 10.1007/BF00297280

Ligase chain reaction (LCR) was evaluated as a tool for the detection of point mutations. For the mutation studied, the specificity of the method is sufficient to detect the mutant allele in the presence of a 200-fold molar excess of the wild-type sequence. LCR was therefore employed in a genetic recombination experiment as a probe for a recessive lethal point mutation. LCR greatly facilitated the isolation of a rare recombinant originating from a crossover event in the 40 kb interval separating the lethal mutation and an enhancer trap insertion in the optomotor-blind locus. The recombinant will allow the study of gene control in situ, in a largely unperturbed regulatory environment.