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Klebsiella pneumoniae  carbapenemase (KPC)-producing  Enterobacterales  are prevalent worldwide and pose an alarming threat to public health. The incidence and transmission of  blaKPC-2  gene via horizontal gene transfer (e.g. transposition) have been well documented. However, the dynamics of transposon structure bearing  blaKPC-2  and their exact effects on the evolution and dissemination of  blaKPC-2  gene are not well characterized. Here, we collected all 161 carbapenem-resistant  Enterobacterales  (CRE) isolates during the early stage of CRE pandemic. We observed that the prevalence of KPC-2-producing  Enterobacterales  was mediated by multiple species and sequence types (STs), and that  blaKPC-2  gene was located on three diverse variants of Tn 1721  in multi-drug resistance (MDR) region of plasmid. Notably, the outbreak of KPC-2-producing plasmid is correlated with the dynamics of transposon structure. Furthermore, we experimentally demonstrated that replicative transposition of Tn 1721  and IS 26  promotes horizontal transfer of  blaKPC-2  and the evolution of KPC-2-producing plasmid. The Tn 1721  variants appearing concurrently with the peak of an epidemic (A2- and B-type) showed higher transposition frequencies and a certain superior ability to propagation. Overall, our work suggests replicative transposition contributes to the evolution and transmission of KPC-2-producing plasmid and highlights its important role in the inter- and intra-species dissemination of  blaKPC-2  gene in  Enterobacterales .

Replicative transposition contributes to the evolution and dissemination of KPC-2-producing plasmid in Enterobacterales