| Summary: | In this study, we found that the deletion of the gene <i>evfG</i> in the type VI secretion system (T6SS) gene cluster significantly affected the motility of porcine extraintestinal pathogenic <i>Escherichia coli</i> (ExPEC) strain PCN033. Furthermore, the bacterial motility assay showed that Δ<i>evfG</i> mutants exhibited reduced motility compared to the parental strain. Transmission electron microscopy (TEM) showed a significant reduction in the number of flagella in the mutant Δ<i>evfG</i> when compared with PCN033. To further explore the reasons why the deletion of <i>evfG</i> affects the motility of PCN033, transcriptomic and metabolomic analyses were conducted. The omics analyses showed that 134 differentially accumulated metabolites and 2236 differentially expressed genes were identified between the mutant Δ<i>evfG</i> and the parental strain PCN033. The metabolome profile and functional annotation analyses indicated that the impaired motility of Δ<i>evfG</i> was connected to the downregulation of the expression levels of genes associated with the energy metabolism pathway and flagellar assembly. Our study provides a new insight into the diminished PCN033 motility induced by <i>evfG</i> deletion. Moreover, the candidate genes and metabolites regulated by the gene <i>evfG</i> in the T6SS, which was involved in the motility of PCN033, were reported in this study.
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