Directed in Vitro Evolution of Therapeutic Bacteriophages: The Appelmans Protocol
The ‘Appelmans protocol’ is used by Eastern European researchers to generate therapeutic phages with novel lytic host ranges. Phage cocktails are iteratively grown on a suite of mostly refractory bacterial isolates until the evolved cocktail can lyse the phage-resistant strains....
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| Format: | Article |
| Language: | English |
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MDPI AG
2019-03-01
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| Series: | Viruses |
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| Online Access: | http://www.mdpi.com/1999-4915/11/3/241 |
| _version_ | 1811210856817491968 |
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| author | Ben H. Burrowes Ian J. Molineux Joe A. Fralick |
| author_facet | Ben H. Burrowes Ian J. Molineux Joe A. Fralick |
| author_sort | Ben H. Burrowes |
| collection | DOAJ |
| description | The ‘Appelmans protocol’ is used by Eastern European researchers to generate therapeutic phages with novel lytic host ranges. Phage cocktails are iteratively grown on a suite of mostly refractory bacterial isolates until the evolved cocktail can lyse the phage-resistant strains. To study this process, we developed a modified protocol using a cocktail of three Pseudomonas phages and a suite of eight phage-resistant (including a common laboratory strain) and two phage-sensitive Pseudomona aeruginosa strains. After 30 rounds of selection, phages were isolated from the evolved cocktail with greatly increased host range. Control experiments with individual phages showed little host-range expansion, and genomic analysis of one of the broad-host-range output phages showed its recombinatorial origin, suggesting that the protocol works predominantly via recombination between phages. The Appelmans protocol may be useful for evolving therapeutic phage cocktails as required from well-defined precursor phages. |
| first_indexed | 2024-04-12T05:02:37Z |
| format | Article |
| id | doaj.art-84f0509ba1cc44e8bdaff8ec08322a69 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-04-12T05:02:37Z |
| publishDate | 2019-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-84f0509ba1cc44e8bdaff8ec08322a692022-12-22T03:46:57ZengMDPI AGViruses1999-49152019-03-0111324110.3390/v11030241v11030241Directed in Vitro Evolution of Therapeutic Bacteriophages: The Appelmans ProtocolBen H. Burrowes0Ian J. Molineux1Joe A. Fralick2Department of Immunology and Molecular Microbiology, Texas Tech University Health Sciences Center, 3601 4th Street, Lubbock, TX 79430, USACenter for Infectious Disease, Department of Molecular Biosciences, The University of Texas at Austin, 1 University Station A5000, Austin, TX 78712, USADepartment of Immunology and Molecular Microbiology, Texas Tech University Health Sciences Center, 3601 4th Street, Lubbock, TX 79430, USAThe ‘Appelmans protocol’ is used by Eastern European researchers to generate therapeutic phages with novel lytic host ranges. Phage cocktails are iteratively grown on a suite of mostly refractory bacterial isolates until the evolved cocktail can lyse the phage-resistant strains. To study this process, we developed a modified protocol using a cocktail of three Pseudomonas phages and a suite of eight phage-resistant (including a common laboratory strain) and two phage-sensitive Pseudomona aeruginosa strains. After 30 rounds of selection, phages were isolated from the evolved cocktail with greatly increased host range. Control experiments with individual phages showed little host-range expansion, and genomic analysis of one of the broad-host-range output phages showed its recombinatorial origin, suggesting that the protocol works predominantly via recombination between phages. The Appelmans protocol may be useful for evolving therapeutic phage cocktails as required from well-defined precursor phages.http://www.mdpi.com/1999-4915/11/3/241Appelmansbacteriophage evolutionbacteriophage recombinationphage therapyPseudomonas aeruginosaantibiotic resistance |
| spellingShingle | Ben H. Burrowes Ian J. Molineux Joe A. Fralick Directed in Vitro Evolution of Therapeutic Bacteriophages: The Appelmans Protocol Viruses Appelmans bacteriophage evolution bacteriophage recombination phage therapy Pseudomonas aeruginosa antibiotic resistance |
| title | Directed in Vitro Evolution of Therapeutic Bacteriophages: The Appelmans Protocol |
| title_full | Directed in Vitro Evolution of Therapeutic Bacteriophages: The Appelmans Protocol |
| title_fullStr | Directed in Vitro Evolution of Therapeutic Bacteriophages: The Appelmans Protocol |
| title_full_unstemmed | Directed in Vitro Evolution of Therapeutic Bacteriophages: The Appelmans Protocol |
| title_short | Directed in Vitro Evolution of Therapeutic Bacteriophages: The Appelmans Protocol |
| title_sort | directed in vitro evolution of therapeutic bacteriophages the appelmans protocol |
| topic | Appelmans bacteriophage evolution bacteriophage recombination phage therapy Pseudomonas aeruginosa antibiotic resistance |
| url | http://www.mdpi.com/1999-4915/11/3/241 |
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