一种非致死提取单头家蝇基因组DNA的简便方法

目的家蝇是一种广泛分布的重要公共卫生害虫,开展其遗传学、分子生物学的研究为认识和利用家蝇、有效地控制蝇传疾病提供科学依据。遗传学、分子生物学的研究常需要用到DNA样品和聚合酶链反应(PCR),建立一种简单快速的DNA提取方法,为研究工作提供极大的便利。方法剪取家蝇的双翅或后足置于提取液中,温育1h后获得家蝇基因组DNA的样品;以此基因组DNA为模板,扩增CYP6D1v1基因;测定去翅或去后足的家蝇繁殖能力。结果基因组DNA成功地从家蝇个体的双翅或后足中提取,此DNA制备液可直接用于PCR扩增;剪除双翅或后足对家蝇个体无致命损伤,不影响家蝇的正常繁殖。结论该方法快速、简便、有效且样品非致死,提取的DNA可以应用于诸如用PCR法鉴定个体基因型等的后续分析。

A simple method for rapid and non lethal isolation of genomic DNA from single flies of Musca domestica

ObjectiveThe house flyMusca domestica(Diptera: Muscidae), is a very important disease vector. Research ingenetics and molecular biology on this pest is helpful for developing an effective control strategy. In genetic studies, it often needsto use the DNA sample and polymerase chain reaction (PCR) technique. A simple and non-lethal method for isolation of genomic DNA should facilitate studies in population genetics and the generation of fly lines of interest.Methods Genomic DNA was isolated from immersing the hind legs or wings of house flies in the lysis buffer and incubating at 37℃for 1 h.Results Genomic DNA was extracted successfully from wings or legs of individual flies. A fragment of the CYP6D1v1 gene was amplified with the DNA as template. Wing or leg ablation did not affect the survival and fertility of flies.Conclusion The method is fast,simple, effective and non-lethal. The DNA prepared by this method can be used directly in PCR for subsequent analysis such as genotyping. Flies without hind legs or wings can produce enough offspring to establish a population.