Synthetic Bacteriophage Technology Helps to Reveal Factors Influencing Reproduction Efficiency of <i>Klebsiella</i> Phages

In this study, synthetic phages with swapped tailspike protein genes were constructed based on Przondovirus bacteriophages KP32_192 and KP32_195, which infect Klebsiella pneumoniae of different capsular serotypes. The specificity of these phages has changed in accordance with the genes of tailspike proteins. However, the reproduction efficiency of these phages strongly depended on the phage scaffold used, the sequence of the N-terminal domain of tail-anchored spikes, and the Klebsiella strain used. Moreover, phages with a non-native scaffold replicated on one of the strains more efficiently than wild-type phage with the same tailspikes. It has also been shown that the sequence of the N-terminal domain of tail-anchored spikes can influence the efficiency of phage adsorption. Therefore, it is necessary to consider not only the capsular specificity of tailspike proteins but also some other genes in the recipient phage genome when designing synthetic phages against the target bacterium.