John Seth G, Mendez Carolina B, Deng Li, Poulos Bonnie, Kauffman Anne Kathryn M, Kern Suzanne, Brum Jennifer, Polz Martin F, Boyle Edward A, Sullivan Matthew B
Environ Microbiol Rep. 2011 Apr;3(2):195-202. doi: 10.1111/j.1758-2229.2010.00208.x.
Ocean viruses alter ecosystems through host mortality, horizontal gene transfer and by facilitating remineralization of limiting nutrients. However, the study of wild viral populations is limited by inefficient and unreliable concentration techniques. Here, we develop a new technique to recover viruses from natural waters using iron-based flocculation and large-pore-size filtration, followed by resuspension of virus-containing precipitates in a pH 6 buffer. Recovered viruses are amenable to gene sequencing, and a variable proportion of phages, depending upon the phage, retain their infectivity when recovered. This Fe-based virus flocculation, filtration and resuspension method (FFR) is efficient (> 90% recovery), reliable, inexpensive and adaptable to many aspects of marine viral ecology and genomics research.
海洋病毒通过宿主死亡、水平基因转移以及促进限制性营养物质的再矿化来改变生态系统。然而,野生病毒种群的研究受到低效且不可靠的浓缩技术的限制。在此,我们开发了一种新技术,利用铁基絮凝和大孔径过滤从天然水体中回收病毒,随后将含病毒的沉淀物重悬于pH 6的缓冲液中。回收的病毒适合进行基因测序,并且根据噬菌体的不同,可变比例的噬菌体在回收时仍保持其感染性。这种基于铁的病毒絮凝、过滤和重悬方法(FFR)高效(回收率>90%)、可靠、廉价且适用于海洋病毒生态学和基因组学研究的许多方面。