Next Article in Journal
Fast PET Scan Tumor Segmentation Using Superpixels, Principal Component Analysis and K-Means Clustering
Next Article in Special Issue
Function of the RNA Coliphage Qβ Proteins in Medical In Vitro Evolution
Previous Article in Journal
Generating CRISPR/Cas9-Derived Mutant Mice by Zygote Cytoplasmic Injection Using an Automatic Microinjector
Article Menu
Issue 1 (March) cover image

Export Article

Open AccessProtocol
Methods Protoc. 2018, 1(1), 6;

Practical Method for Isolation of Phage Deletion Mutants

Instituto de Productos Lácteos de Asturias (IPLA-CSIC), Paseo Río Linares s/n, 33300 Villaviciosa, Spain
Author to whom correspondence should be addressed.
Received: 12 December 2017 / Revised: 9 January 2018 / Accepted: 15 January 2018 / Published: 17 January 2018
Full-Text   |   PDF [4502 KB, uploaded 18 January 2018]   |  


The growing concern about multi-drug resistant pathogenic bacteria has led to a renewed interest in the study of bacteriophages as antimicrobials and as therapeutic agents against infectious diseases (phage therapy). Phages to be used for this purpose have to be subjected to in-depth genomic characterization. It is essential to ascribe specific functions to phage genes, which will give information to unravel phage biology and to ensure the lack of undesirable genes, such as virulence and antibiotic resistance genes. Here, we describe a simple protocol for the selection of phage mutants carrying random deletions along the phage genome. Theoretically, any DNA region might be removed with the only requirement that the phage particle viability remains unaffected. This technique is based on the instability of phage particles in the presence of chelating compounds. A fraction of the phage population naturally lacking DNA segments will survive the treatment. Within the context of phages as antimicrobials, this protocol is useful to select lytic variants from temperate phages. In terms of phage efficiency, virulent phages are preferred over temperate ones to remove undesirable bacteria. This protocol has been used to obtain gene mutations that are involved in the lysogenic cycle of phages infecting Gram-positive bacteria (Staphylococcus and Lactobacillus). View Full-Text
Keywords: phage deletion mutant; chelating agent; temperate phage; virulent phage phage deletion mutant; chelating agent; temperate phage; virulent phage

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

Share & Cite This Article

MDPI and ACS Style

Gutiérrez, D.; Fernández, L.; Rodríguez, A.; García, P. Practical Method for Isolation of Phage Deletion Mutants. Methods Protoc. 2018, 1, 6.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Metrics

Article Access Statistics



[Return to top]
Methods Protoc. EISSN 2409-9279 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top