UNIT 13.3 Isolation of Bacteriophages from Environmental Sources, and Creation and Functional Screening of Phage DNA Libraries. Issue 7 (23rd October 2013)

• Record Type: Journal Article
• Authors: Pelzek, Adam J.; Schuch, Raymond; Schmitz, Jonathan E.; Fischetti, Vincent A.
• Is Part Of: Current protocols essential laboratory techniques. Issue 7(2014)
• Page Start: 13.3.1
• Page End: 13.3.35
• Publication Date: 2013-10-23
• Subjects: bacteriophage -- phage -- phage isolation -- phage culture -- metagenomics -- phage titer -- phage characterization -- phage DNA -- single‐step growth curve -- DNA library -- lysin screen; Life sciences -- Laboratory manuals; Medical laboratories; Diagnosis, Laboratory; Clinical Laboratory Techniques; Clinical Protocols; Diagnosis, Laboratory; Medical laboratories; 570.28
• DOI: 10.1002/9780470089941.et1303s07
• Languages: English
• ISSNs: 1948-3430

Abstract

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.