膜过滤固定化技术分离和富集噬菌体

Q3 Biochemistry, Genetics and Molecular Biology Current Protocols in Cell Biology Pub Date : 2018-05-18 DOI:10.1002/cpcb.41
Gaurav S. Ghugare, Vijay D. Nimkande, Krishna Khairnar
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引用次数: 1

摘要

本文描述的方法能够从环境样品中快速分离和富集宿主特异性噬菌体。这是通过使用一个简单的0.45µm毫孔膜来实现的,其中特定的宿主固定在膜上,并在膜过滤装置的帮助下将怀疑含有噬菌体的样品暴露在固定的细胞中。这一过滤步骤促进了噬菌体与宿主的特异性相互作用,并利用经典的膜过滤方法使这种相互作用最大化。在真空泵的真空作用下,过滤器组件为样品中的每个噬菌体提供了与膜过滤器上的特定宿主相互作用的机会。我们的技术可以通过吸附将特定的噬菌体与其宿主细胞一起保留在膜上;这些吸附在滤盘上的噬菌体(连同它们的宿主),然后通过培养在常规营养肉汤——色氨酸豆汤(TSB)中进行富集。借助空斑测定方法,可以分离、分离和富集各种细菌的宿主特异性噬菌体。©2018 by John Wiley &儿子,Inc。
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Isolation and Enrichment of Bacteriophages by Membrane Filtration Immobilization Technique

The method described here enables rapid bacteriophage isolation and enrichment of host-specific bacteriophages from an environmental sample. This is achieved by using a simple 0.45-µm Millipore membrane where a specific host is immobilized on the membrane and a sample suspected of containing bacteriophages is exposed to the immobilized cells with the help of a membrane filtration unit. This filtration step facilitates host-specific interaction of bacteriophages with the host and maximization of this interaction using a classic membrane filtration method. Under the effect of vacuum from a vacuum pump, a filter assembly provides a chance for every bacteriophage in the sample to interact with the specific host on the membrane filter. Our technique allows retaining specific bacteriophages on the membrane along with its host cells via adsorption; these adsorbed bacteriophages (along with their hosts) on a filter disc are then enriched in regular nutritive broth, tryptone soya broth (TSB), by incubation. With help of a plaque assay method, host-specific phages of various bacterial species can be isolated, segregated, and enriched. © 2018 by John Wiley & Sons, Inc.

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Current Protocols in Cell Biology
Current Protocols in Cell Biology Biochemistry, Genetics and Molecular Biology-Cell Biology
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期刊介绍: Developed by leading scientists in the field, Current Protocols in Cell Biology is an essential reference for researchers who study the relationship between specific molecules and genes and their location, function and structure at the cellular level. Updated every three months in all formats, CPCB is constantly evolving to keep pace with the very latest discoveries and developments.
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