Identification of EMS-Induced Mutations inDrosophila melanogasterby Whole-Genome Sequencing
- 1 May 2009
- journal article
- Published by Oxford University Press (OUP) in Genetics
- Vol. 182 (1), 25-32
- https://doi.org/10.1534/genetics.109.101998
Abstract
Next-generation methods for rapid whole-genome sequencing enable the identification of single-base-pair mutations in Drosophila by comparing a chromosome bearing a new mutation to the unmutagenized sequence. To validate this approach, we sought to identify the molecular lesion responsible for a recessive EMS-induced mutation affecting egg shell morphology by using Illumina next-generation sequencing. After obtaining sufficient sequence from larvae that were homozygous for either wild-type or mutant chromosomes, we obtained high-quality reads for base pairs composing ∼70% of the third chromosome of both DNA samples. We verified 103 single-base-pair changes between the two chromosomes. Nine changes were nonsynonymous mutations and two were nonsense mutations. One nonsense mutation was in a gene, encore, whose mutations produce an egg shell phenotype also observed in progeny of homozygous mutant mothers. Complementation analysis revealed that the chromosome carried a new functional allele of encore, demonstrating that one round of next-generation sequencing can identify the causative lesion for a phenotype of interest. This new method of whole-genome sequencing represents great promise for mutant mapping in flies, potentially replacing conventional methods.This publication has 20 references indexed in Scilit:
- Rapid whole-genome mutational profiling using next-generation sequencing technologiesGenome Research, 2008
- Retention of Induced Mutations in a Drosophila Reverse-Genetic ResourceGenetics, 2008
- Mapping short DNA sequencing reads and calling variants using mapping quality scoresGenome Research, 2008
- Caenorhabditis elegans mutant allele identification by whole-genome sequencingNature Methods, 2008
- A high-resolution, nucleosome position map of C. elegans reveals a lack of universal sequence-dictated positioningGenome Research, 2008
- High-Resolution Profiling of Histone Methylations in the Human GenomeCell, 2007
- All Paired Up with No Place to Go: Pairing, Synapsis, and DSB Formation in a Balancer HeterozygotePLoS Genetics, 2005
- Genome sequencing in microfabricated high-density picolitre reactorsNature, 2005
- Target-selected mutant screen by TILLING in DrosophilaGenome Research, 2005
- Encore facilitates SCF-Ubiquitin-proteasome-dependent proteolysis duringDrosophilaoogenesisDevelopment, 2003