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
Laboratory of Molecular Microbiology, Institute of Chemical Biology and Fundamental Medicine of the Siberian Branch of the Russian Academy of Sciences, Novosibirsk 630090, Russia.
Shared Research Facility "Siberian Circular Photon Source" (SRF "SKIF"), Boreskov Institute of Catalysis of the Siberian Branch of the Russian Academy of Sciences, Novosibirsk 630090, Russia.
Int J Mol Sci. 2025 Jul 16;26(14):6824. doi: 10.3390/ijms26146824.
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 phages, KP192 and KP195, which infect 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 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 (). However, the efficiency of plating and the lytic efficiency of these phages strongly depended on the genomic scaffold used and the 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 gene transplantation can have a profound impact on the efficiency of phage propagation on target bacterial strains.
在本研究中，通过在两种感染不同荚膜类型的噬菌体KP192和KP195之间交换围绕尾刺基因的基因组支架，研究了基因组支架对噬菌体特性的影响。采用基于酵母的转化相关重组克隆技术以及随后在细菌中对合成噬菌体基因组进行“重启”来构建噬菌体。使用具有K2、K64和KL111荚膜类型的菌株，结果表明合成噬菌体的荚膜特异性与尾刺蛋白的荚膜特异性完全一致（）。然而，这些噬菌体的平板接种效率和裂解效率强烈依赖于所使用的基因组支架和所使用的菌株。与野生型噬菌体相比，基因组支架交换后的合成噬菌体在多种菌株上表现出更高的繁殖效率，这表明交换后的基因组支架的某些元件可提高噬菌体复制效率，可能是通过阻断宿主的一些抗噬菌体防御系统来实现的。我们的研究结果表明，即使对于亲缘关系密切的噬菌体，用于基因移植的基因组支架的选择也会对噬菌体在目标细菌菌株上的繁殖效率产生深远影响。
