Rapid identification of heterozygous mutations in Drosophila melanogaster using genomic capture sequencing
作者： H. Wang , A. Chattopadhyay , Z. Li , B. Daines , Y. Li
关键词: Biology 、 Mutation (genetic algorithm) 、 Genome 、 Mutant 、 Drosophila melanogaster 、 Genetics 、 Point mutation 、 Forward genetics 、 Gene mapping 、 Genetic model
摘要: One of the key advantages using Drosophila melanogaster as a genetic model organism is ability to conduct saturation mutagenesis screens identify genes and pathways underlying given phenotype. Despite large number tools developed facilitate downstream cloning mutations obtained from such screens, current procedure remains labor intensive, time consuming, costly. To address this issue, we designed an efficient strategy for rapid identification heterozygous in fly genome by combining rough mapping, targeted DNA capture, second generation sequencing technology. We first tested method on flies carrying either previously characterized dac5 or sensE2 mutation. Targeted amplification genomic regions near these two loci was used enrich sequencing, both point were successfully identified. When applied uncharacterized twr mutant flies, mutation identified single-base gene Spase18-21. This targeted-genome-sequencing reduces effort required up 80% compared with approach lowers cost <$1000 each mutant. Introduction other sequencing-based methods will enable broader usage forward genetics have significant impacts field organisms Drosophila.
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