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首页> 外文期刊>Current Protocols in Cell Biology >Isolation and Enrichmentof Bacteriophages by MembraneFiltration Immobilization Technique
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Isolation and Enrichmentof Bacteriophages by MembraneFiltration Immobilization Technique

机译：膜异常固定技术分离和富集噬菌体

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摘要

The method described here enables rapid bacteriophage isolation and enrichmentof host-specific bacteriophages from an environmental sample. This isachieved by using a simple 0.45-μm Millipore membrane where a specific hostis immobilized on the membrane and a sample suspected of containing bacteriophagesis exposed to the immobilized cells with the help of a membranefiltration unit. This filtration step facilitates host-specific interaction of bacteriophageswith the host and maximization of this interaction using a classicmembrane filtration method. Under the effect of vacuum from a vacuum pump,a filter assembly provides a chance for every bacteriophage in the sample tointeract with the specific host on the membrane filter. Our technique allowsretaining specific bacteriophages on the membrane along with its host cells viaadsorption; these adsorbed bacteriophages (along with their hosts) on a filterdisc are then enriched in regular nutritive broth, tryptone soya broth (TSB),by incubation. With help of a plaque assay method, host-specific phages ofvarious bacterial species can be isolated, segregated, and enriched.
机译：这里描述的方法能够从环境样品中快速噬菌体分离和富集宿主特异性噬菌体。这种通过使用简单的0.45μmmillipore膜来解析，其中在膜上固定在膜上的特定主体和涉及含有膜的含有暴露于固定细胞的噬菌体的样品。该过滤步骤促进了宿主的宿主特异性相互作用，并使用分类模糊过滤方法最大化该相互作用。在真空泵的真空的影响下，过滤器组件为样品中的每种噬菌体提供了与膜过滤器上的特定主体的每个噬菌体的机会。我们的技术使膜上的特异性噬菌体以及其宿主细胞的负载率脱发;然后通过孵育，将这些吸附的噬菌体（以及它们的宿主及其宿主）富含常规营养肉汤，培养醇大豆肉汤（TSB）。借助斑块测定方法，可以分离，分离和富集的宿主特异性噬菌体。

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  • 来源
    《Current Protocols in Cell Biology》 |2018年第1期|共8页
  • 作者

    Gaurav S. Ghugare; Vijay D. Nimkande; Krishna Khairnar;

  • 作者单位

    Environmental Virology Cell Council of Scientific and Industrial Research—NationalEnvironmental Engineering Research Institute (CSIR NEERI) Nehru Marg Nagpur Maharashtra India;

    Environmental Virology Cell Council of Scientific and Industrial Research—NationalEnvironmental Engineering Research Institute (CSIR NEERI) Nehru Marg Nagpur Maharashtra India;

    Environmental Virology Cell Council of Scientific and Industrial Research—NationalEnvironmental Engineering Research Institute (CSIR NEERI) Nehru Marg Nagpur Maharashtra India;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 细胞生物学;
  • 关键词

    bacteriophages enrichment; membrane; plaque assay;

    机译：噬菌体富集;膜;斑块测定;

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