Replacement of the Genomic Scaffold Improves the Replication Efficiency of Synthetic Klebsiella Phages

In this study, the impact of the genomic scaffold on the properties of bacteriophages was investigated by swapping the genomic scaffolds surrounding the tailspike genes between two Przondovirus phages, KP192 and KP195, which infect Klebsiella pneumoniae with different capsular types. A yeast-based t...

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Bibliographic Details
Published in:International journal of molecular sciences Vol. 26; no. 14; p. 6824
Main Authors: Baykov, Ivan K., Kurchenko, Olga M., Mikhaylova, Ekaterina E., Miroshnikova, Anna V., Morozova, Vera V., Khlebnikova, Marianna I., Tikunov, Artem Yu, Kozlova, Yuliya N., Tikunova, Nina V.
Format: Journal Article
Language:English
Published: Switzerland MDPI AG 16.07.2025
MDPI
Subjects:
Amino acids
Bacterial pneumonia
Bacteriophages - genetics
Bacteriophages - physiology
Biology
Cloning
Efficiency
Genes
Genome, Viral
Genomes
Genomics
Klebsiella pneumoniae - genetics
Klebsiella pneumoniae - virology
Phages
Pneumonia
Proteins
Viral Tail Proteins - genetics
Virus Replication - genetics
United States
Massachusetts
Russia
Klebsiella
synthetic biology
receptor-binding protein
transformation-associated recombination cloning
tailspike depolymerase
bacteriophage
genome assembly
ISSN:1422-0067, 1661-6596, 1422-0067
Online Access:Get full text
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Summary:In this study, the impact of the genomic scaffold on the properties of bacteriophages was investigated by swapping the genomic scaffolds surrounding the tailspike genes between two Przondovirus phages, KP192 and KP195, which infect Klebsiella pneumoniae with different capsular types. A yeast-based transformation-associated recombination cloning technique and subsequent “rebooting” of synthetic phage genomes in bacteria were used to construct the phages. Using Klebsiella strains with K2, K64, and KL111 capsular types, it was shown that the capsular specificity of the synthetic phages is fully consistent with that of the tailspike proteins (tsp). However, the efficiency of plating and the lytic efficiency of these phages strongly depended on the genomic scaffold used and the Klebsiella strain used. Synthetic phages with swapped genomic scaffolds demonstrated superior reproduction efficiency using a number of strains compared to wild-type phages, indicating that some elements of the swapped genomic scaffold enhance phage replication efficiency, presumably by blocking some of the host anti-phage defense systems. Our findings demonstrate that even in the case of closely related phages, the selection of the genomic scaffold used for tsp gene transplantation can have a profound impact on the efficiency of phage propagation on target bacterial strains.
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ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms26146824