Unit

UNIT 13.3 Isolation of Bacteriophages from Environmental Sources, and Creation and Functional Screening of Phage DNA Libraries

  1. Adam J. Pelzek1,
  2. Raymond Schuch2,
  3. Jonathan E. Schmitz3,
  4. Vincent A. Fischetti4

Published Online: 23 OCT 2013

DOI: 10.1002/9780470089941.et1303s07

Current Protocols Essential Laboratory Techniques

Current Protocols Essential Laboratory Techniques

How to Cite

Pelzek, A. J., Schuch, R., Schmitz, J. E. and Fischetti, V. A. 2013. Isolation of Bacteriophages from Environmental Sources, and Creation and Functional Screening of Phage DNA Libraries. Current Protocols Essential Laboratory Techniques. 13:13.3:13.3.1–13.3.35.

Author Information

  1. 1

    New York University School of Medicine, New York, New York

  2. 2

    ContraFect Corporation, Yonkers, New York

  3. 3

    Vanderbilt University Medical Center, Nashville, Tennessee

  4. 4

    Vincent A. Fischetti Laboratory of Bacterial Pathogenesis and Immunology, The Rockefeller University, New York, New York

Publication History

  1. Published Online: 23 OCT 2013

Abstract

Bacteriophages (phages) have evolved specific classes of proteins such as adhesins and lysins to mediate specific bacterial host recognition and rapid and efficient lysis following infection. However, because many bacterial species, and thus phages, cannot be cultured in the laboratory, techniques for the efficient cloning and screening of phage gene libraries are required to aid in the discovery of novel phage proteins for use in diagnostic and therapeutic applications. This article contains protocols for the isolation of phages from environmental samples, enrichment of phages targeting host bacteria of interest, and induction of phages from lysogenized host strains. We also provide an optimized protocol for the creation and functional screening of phage DNA libraries derived from environmental samples. Curr. Protoc. Essential Lab. Tech. 7:13.3.1-13.3.35. © 2013 by John Wiley & Sons, Inc.

Keywords:

  • bacteriophage;
  • phage;
  • phage isolation;
  • phage culture;
  • metagenomics;
  • phage titer;
  • phage characterization;
  • phage DNA;
  • single-step growth curve;
  • DNA library;
  • lysin screen