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Analysing parallel strategies to alter the host specificity of bacteriophage T7
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Avramucz, Akos, Møller-Olsen, Christian, Grigonyte, Aurelija M., Paramalingam, Yanahan, Millard, Andrew, Sagona, Antonia P. and Feher, Tamas (2021) Analysing parallel strategies to alter the host specificity of bacteriophage T7. Biology, 10 (6). 556. doi:10.3390/biology10060556 ISSN 2079-7737.
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Official URL: https://doi.org/10.3390/biology10060556
Abstract
The recognition and binding of host bacteria by bacteriophages is most often enabled by a highly specific receptor–ligand type of interaction, with the receptor-binding proteins (RBPs) of phages being the primary determinants of host specificity. Specifically modifying the RBPs could alter or extend the host range of phages otherwise exhibiting desired phenotypic properties. This study employed two different strategies to reprogram T7 phages ordinarily infecting commensal K12 Escherichia coli strains to infect pathogen-associated K1-capsule-expressing strains. The strategies were based on either plasmid-based homologous recombination or bacteriophage recombineering using electroporated DNA (BRED). Our work pursued the construction of two genetic designs: one replacing the gp17 gene of T7, the other replacing gp11, gp12, and gp17 of T7 with their K1F counterparts. Both strategies displayed successful integration of the K1F sequences into the T7 genome, detected by PCR screening. Multiple methods were utilised to select or enrich for chimeric phages incorporating the K1F gp17 alone, including trxA, host-specificity, and CRISPR-Cas-based selection. Irrespective of the selection method, the above strategy yielded poorly reproducible phage propagation on the new host, indicating that the chimeric phage was less fit than the wild type and could not promote continual autonomous reproduction. Chimeric phages obtained from BRED incorporating gp11-12 and gp17, however, all displayed infection in a 2-stage pattern, indicating the presence of both K1F and T7 phenotypes. This study shows that BRED can be used as a tool to quickly access the potential of new RBP constructs without the need to engineer sustainably replicating phages. Additionally, we show that solely repurposing the primary RBP is, in some cases, insufficient to produce a viable chimeric phage. View Full-Text
Item Type: | Journal Article | ||||||||||||
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Subjects: | Q Science > QR Microbiology > QR355 Virology | ||||||||||||
Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||||||||||
Library of Congress Subject Headings (LCSH): | Bacteriophages, Antibiotics, Drug resistance in microorganisms, Proteins | ||||||||||||
Journal or Publication Title: | Biology | ||||||||||||
Publisher: | M D P I AG | ||||||||||||
ISSN: | 2079-7737 | ||||||||||||
Official Date: | 20 June 2021 | ||||||||||||
Dates: |
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Volume: | 10 | ||||||||||||
Number: | 6 | ||||||||||||
Article Number: | 556 | ||||||||||||
DOI: | 10.3390/biology10060556 | ||||||||||||
Status: | Peer Reviewed | ||||||||||||
Publication Status: | Published | ||||||||||||
Access rights to Published version: | Open Access (Creative Commons) | ||||||||||||
Date of first compliant deposit: | 16 June 2021 | ||||||||||||
Date of first compliant Open Access: | 21 June 2021 | ||||||||||||
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