Colombet J, Robin A, Lavie L, Bettarel Y, Cauchie H M, Sime-Ngando T
Laboratoire de Biologie des Protistes, Université Blaise Pascal (Clermont-Ferrand II), UMR CNRS 6023, F-63177, Aubière Cedex, France.
J Microbiol Methods. 2007 Dec;71(3):212-9. doi: 10.1016/j.mimet.2007.08.012. Epub 2007 Sep 4.
We have described the use of Polyethylene glycol (PEG) for the precipitation of natural communities of aquatic viruses, and its comparison with the usual concentration method based on ultracentrifugation. Experimental samples were obtained from different freshwater ecosystems whose trophic status varied. Based on transmission electron microscope observations and counting of phage-shaped particles, our results showed that the greatest recovery efficiency for all ecosystems was obtained when we used the PEG protocol. On average, this protocol allowed the recovery of >2-fold more viruses, compared to ultracentrifugation. In addition, the diversity of virioplankton, based on genomic size profiling using pulsed field gel electrophoresis, was higher and better discriminated when we used the PEG method. We conclude that pegylation offers a valid, simple and cheaper alternative method to ultracentrifugation, for the concentration and the purification of pelagic viruses.
我们已经描述了使用聚乙二醇(PEG)沉淀水生病毒自然群落的方法,并将其与基于超速离心的常规浓缩方法进行了比较。实验样本取自营养状态不同的不同淡水生态系统。基于透射电子显微镜观察和噬菌体形状颗粒的计数,我们的结果表明,当使用PEG方案时,所有生态系统的回收效率最高。平均而言,与超速离心相比,该方案能回收的病毒数量多出两倍以上。此外,当使用PEG方法时,基于脉冲场凝胶电泳的基因组大小分析,浮游病毒的多样性更高且区分效果更好。我们得出结论,聚乙二醇化提供了一种有效、简单且成本更低的替代超速离心的方法,用于浓缩和纯化浮游病毒。