Pausz Clemens, Clasen Jessica L, Suttle Curtis A
Department of Earth & Ocean Sciences, University of British Columbia, Vancouver, BC, Canada.
Methods Mol Biol. 2009;502:255-78. doi: 10.1007/978-1-60327-565-1_15.
Since most of the phage genomes isolated from natural samples are previously unknown sequences, an isolation-independent approach is necessary to quantify the diversity of natural viral communities. Currently, two different methodological approaches are widely used to obtain genetic fingerprints of natural phage communities. While the separation of different viral genomes with pulsed field gel electrophoresis (PFGE) is based on the size of the genome, denaturing gradient gel electrophoresis (DGGE) uses minor differences in gene base composition to separate fragments of amplified DNA from natural viral communities. Finger printing techniques are a relatively fast and cheap tool to assess the diversity of environmental viruses. Together, PFGE and DGGE provide useful tools to study viral ecology in natural habitats.
由于从自然样本中分离出的大多数噬菌体基因组都是以前未知的序列,因此需要一种不依赖分离的方法来量化天然病毒群落的多样性。目前,两种不同的方法被广泛用于获取天然噬菌体群落的遗传指纹。虽然用脉冲场凝胶电泳(PFGE)分离不同的病毒基因组是基于基因组的大小,但变性梯度凝胶电泳(DGGE)利用基因碱基组成的微小差异来分离天然病毒群落中扩增DNA的片段。指纹技术是评估环境病毒多样性的一种相对快速且廉价的工具。PFGE和DGGE共同为研究自然栖息地中的病毒生态学提供了有用的工具。