A fluorescent quantitative PCR approach to map gene deletions in the Drosophila genome.
We report the application of TaqMan quantitative PCR (QPCR) to map Drosophila chromosome deficiencies by discrimination of twofold copy number differences. For a model system, we used this technology to confirm the X chromosomal mapping of Dspt6 given the autosomal mapping of Dspt4. We then used this technique on both preexisting deletion mutant flies and flies that we generated with deletions to demonstrate the presence or absence of Dspt6, Dspt4, and swa in various deletion mutant flies. In contrast with in situ hybridization studies, QPCR both vitiates the need to do these more intricate studies, and it is more accurate as the site of deletion can be known down to the 10(2)-bp level. We then successfully applied the technique to the analysis of transcription, demonstrating that the amount of Dspt6 or Dspt4 transcriptional product depended directly on the dosage of the Dspt6 or Dspt4 gene, respectively. The rapidity and precision of this method demonstrates its applicability in Drosophila genetics, the rapid and accurate mapping of Drosophila deletion mutants.
Chiang
P W
PW
Department of Pediatrics, University of Michigan, Ann Arbor, Michigan 48109-0650, USA. pwchiang@umich.edu
Wei
W L
WL
Gibson
K
K
Bodmer
R
R
Kurnit
D M
DM
D000818:Animals; D002874:Chromosome Mapping; D003433:Crosses, Genetic; D004331:Drosophila melanogaster / Q000235:genetics*; D005260:Female; D005298:Fertility / Q000235:genetics; D017353:Gene Deletion; D005819:Genetic Markers; D016678:Genome; D008297:Male; D016296:Mutagenesis; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics; D013050:Spectrometry, Fluorescence; D014158:Transcription, Genetic; D014960:X Chromosome
D005819:Genetic Markers; D012333:RNA, Messenger