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使用聚合酶链反应的随机引物在DNA水平上解析地中海实蝇种群。

Resolution of populations of the Mediterranean fruit fly at the DNA level using random primers for the polymerase chain reaction.

作者信息

Haymer D S, McInnis D O

机构信息

Department of Genetics and Molecular Biology, University of Hawaii, Honolulu 96822.

出版信息

Genome. 1994 Apr;37(2):244-8. doi: 10.1139/g94-034.

Abstract

We have used the polymerase chain reaction (PCR) and the random amplified polymorphic DNA (RAPD) method to identify DNA polymorphisms that can be used as genetic markers to characterize populations of the Mediterranean fruit fly, Ceratitis capitata. In this study, RAPD markers have been used to resolve genetic variability between populations of this major agricultural pest species. The populations analyzed represent either laboratory stocks or wild collections originating from different geographic localities. Using the same set of individual flies from each of several populations, we show that the use of different primers in the RAPD method permits detection of different levels of population differentiation. We show results from RAPD primers (e.g., primer 14) that identify regions of the genome (through PCR amplification) that are essentially monomorphic in all flies originating from a particular geographic locality. We also show RAPD primers (e.g., primer 67) that identify what appear to be highly variable regions of the genome. We have used primers of this type to produce genetic markers that can distinguish even between laboratory versus wild populations as well as subpopulations of flies from more broadly defined geographic localities, such as within the Hawaiian islands. These results show that the RAPD method is a broadly applicable, high resolution method for documenting genetic variability within and between populations of insect pest species.

摘要

我们已使用聚合酶链反应(PCR)和随机扩增多态性DNA(RAPD)方法来鉴定DNA多态性,这些多态性可作为遗传标记,用于表征地中海实蝇(Ceratitis capitata)的种群。在本研究中,RAPD标记已被用于解析这种主要农业害虫种群之间的遗传变异性。所分析的种群要么是实验室品系,要么是来自不同地理区域的野生采集样本。使用来自几个种群中每个种群的同一组个体果蝇,我们表明在RAPD方法中使用不同的引物能够检测到不同水平的种群分化。我们展示了RAPD引物(例如引物14)的结果,这些引物通过PCR扩增识别出在源自特定地理区域的所有果蝇中基本为单态的基因组区域。我们还展示了RAPD引物(例如引物67),这些引物识别出似乎是基因组中高度可变的区域。我们已使用这类引物来产生遗传标记,这些标记甚至能够区分实验室种群与野生种群,以及来自更广泛定义地理区域(如夏威夷群岛内部)的果蝇亚种群。这些结果表明,RAPD方法是一种广泛适用的、高分辨率的方法,用于记录害虫种群内部和种群之间的遗传变异性。

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