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利用变性梯度凝胶电泳(DGGE)对寄生虫核糖体DNA中的指纹序列变异进行分析

Fingerprinting sequence variation in ribosomal DNA of parasites by DGGE.

作者信息

Gasser R, Nansen P, Guldberg P

机构信息

University of Melbourne, Department of Veterinary Science, Werribee, Australia.

出版信息

Mol Cell Probes. 1996 Apr;10(2):99-105. doi: 10.1006/mcpr.1996.0014.

Abstract

Although there is a tendency for rDNA genes within a species to maintain sequence homogeneity, there can be significant levels of variation among rDNA repeat sequences within populations or individuals of a species as a consequence of mutation mechanisms. To date, there have been no practical techniques available in molecular parasitology that allow the extent of sequence variation among the repeats (i.e., number of sequence types) to be displayed visually. In this report, we describe the use of the denaturing gradient gel electrophoresis (DGGE) technique for the rapid screening of parasite rDNA for sequence variation without the need for exhaustive cloning or DNA sequencing. The resolution of this variation by DGGE provides a diagnostic fingerprint for a species.

摘要

尽管一个物种内的核糖体DNA(rDNA)基因有保持序列同质性的趋势,但由于突变机制,一个物种的种群或个体内的rDNA重复序列之间可能存在显著水平的变异。迄今为止,分子寄生虫学中尚无实用技术能够直观显示重复序列之间的序列变异程度(即序列类型数量)。在本报告中,我们描述了使用变性梯度凝胶电泳(DGGE)技术快速筛选寄生虫rDNA的序列变异,而无需进行详尽的克隆或DNA测序。通过DGGE对这种变异的分辨率为一个物种提供了诊断指纹。

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