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通过线粒体DNA的PCR-RFLP揭示的蝇类多样性

Fly diversity revealed by PCR-RFLP of mitochondrial DNA.

作者信息

Asraoui Jimmy F, Sayar Nancy P, Knio Khouzama M, Smith Colin A

机构信息

Department of Biology, American University of Beirut, Beirut, Lebanon.

出版信息

Biochem Mol Biol Educ. 2008 Sep;36(5):354-62. doi: 10.1002/bmb.20220.

Abstract

In this article, we describe an inexpensive, two-session undergraduate laboratory activity that introduces important molecular biology methods in the context of biodiversity. In the first session, students bring tentatively identified flies (order Diptera, true flies) to the laboratory, extract DNA, and amplify a region of the mitochondrial gene NADH dehydrogenase subunit 1. In the second session, the students digest the PCR product with a restriction enzyme, visualize the resulting fragments by agarose gel electrophoresis, and analyze their results with comparison to known sequences. The diversity of flies and their importance as disease vectors, agriculture pests, pollinators, models of speciation, and in the case of Drosophila melanogaster, as a genetic model organism, offer many perspectives with which to appeal to students' interests. The laboratory exercise can be linked as a module to topics in biodiversity, bioinformatics, entomology, evolution, and mutagenesis.

摘要

在本文中,我们描述了一项成本低廉、分两个阶段进行的本科实验室活动,该活动在生物多样性背景下介绍重要的分子生物学方法。在第一阶段,学生们将初步鉴定的苍蝇(双翅目,实蝇)带到实验室,提取DNA,并扩增线粒体基因烟酰胺腺嘌呤二核苷酸脱氢酶亚基1的一个区域。在第二阶段,学生们用一种限制性酶消化PCR产物,通过琼脂糖凝胶电泳观察所得片段,并与已知序列进行比较分析结果。苍蝇的多样性及其作为疾病传播媒介、农业害虫、传粉者、物种形成模型以及(以黑腹果蝇为例)作为遗传模式生物的重要性,提供了许多能吸引学生兴趣的视角。该实验室练习可以作为一个模块与生物多样性、生物信息学、昆虫学、进化和诱变等主题相联系。

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