Identification of EMS-induced mutations in Drosophila melanogaster by whole-genome sequencing.
Genetics
; 182(1): 25-32, 2009 May.
Article
en En
| MEDLINE
| ID: mdl-19307605
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 approximately 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.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Genoma
/
Drosophila melanogaster
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Metanosulfonato de Etilo
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Estudio de Asociación del Genoma Completo
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Mutágenos
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Mutación
Tipo de estudio:
Diagnostic_studies
Límite:
Animals
Idioma:
En
Revista:
Genetics
Año:
2009
Tipo del documento:
Article
País de afiliación:
Estados Unidos