<p>Comparative analysis of KPC-2-encoding chimera plasmids with multi-replicon IncR:Inc_pA1763-KPC:IncN1 or IncFII_pHN7A8:Inc_pA1763-KPC: IncN1</p>

Abstract

BackgroundIncR, IncFII, IncpA1763-KPC, and IncN1 plasmids have been increasingly found among Enterobacteriaceae species, but plasmids with hybrid structures derived from the above-mentioned incompatibility groups have not yet been described.MethodsPlasmids p721005-KPC, p504051-KPC, and pA3295-KPC were fully sequenced and compared with previously sequenced related plasmids pHN84KPC (IncR), pKPHS2 (IncFIIK), pKOX_NDM1 (IncFIIY), pHN7A8 (IncFIIpHN7A8), and R46 (IncN1).ResultsThe backbone of p721005-KPC/p504051-KPC was a hybrid of the entire 10-kb IncR-type backbone from pHN84KPC, the entire 64.3-kb IncFIIK-type maintenance, and conjugal transfer regions from pKPHS2, a 15.5-kb IncFIIY-type maintenance region from pKOX_NDM1 and a 5.6-kb IncpA1763-KPC-type backbone region from pA1763-KPC, and it contained a primary IncR replicon and two auxiliary IncpA1763-KPC and IncN1 replicons. The backbone of pA3295-KPC was a hybrid of a 7.2-kb IncFIIpHN7A8-type backbone region from pHN7A8, the almost entire 33.3-kb IncN1-type maintenance and conjugal transfer regions highly similar to R46, a 26.2-kb IncFIIK-type maintenance regions from pKPHS2, the above 15.5-kb IncFIIY-type maintenance region, and the above 5.6-kb IncpA1763-KPC-type backbone region, and it contained a primary Inc-FIIpHN7A8 replicon and two auxiliary IncpA1763-KPC and IncN1 replicons. Each of p721005-KPC, p504051-KPC, and pA3295-KPC acquired a wealth of accessory modules, carrying a range of intact and residue mobile elements (such as insertion sequences, unit transposons, and integrons) and resistance markers (such as blaKPC, tetA, dfrA, and qnr).ConclusionIn each of p721005-KPC, p504051-KPC, and pA3295-KPC, multiple replicons in coordination with maintenance and conjugation regions of various origins would maintain a broad host range and a stable replication at a steady-state plasmid copy number.

<p>Complete-Genome Sequencing and Comparative Genomic Characterization of an IMP-4 Producing <em>Citrobacter freundii</em> Isolate from Patient with Diarrhea</p>

“…In addition, the plasmid carries multiple mobile transfer elements, which plays an important role in the propagation of the plasmid. 44 The backbone of IncN1 type plasmid includes regions of replication, maintenance, and conjugal transfer. Certain plasmid types have been implicated in the endemic spread of IMP-4 in Australia.…”

Characterization of an IncR Plasmid with Two Copies of ISCR-Linked qnrB6 from ST968 Klebsiella pneumoniae

“…Plasmid pHN84KPC (KY296104) was selected as reference because it contains the most complete IncR backbone, and gene sequences with coverage > 50% and identity > 80% to those of pHN84KPC were considered effective homologous backbone genes [ 5 ]. Sequences containing the backbone genes resD , repB , parAB , and umuCD were used for the phylogenetic analysis.…”

“…The first sequenced IncR single-replicon plasmid was pEFER, though IncR often coexists with other replicons such as IncC, IncN, IncHI, and IncFII [ 2 ]. There are an increasing number of reports about IncR plasmids carrying various resistance genes, such as bla KPC-2 , bla DHA-1 , bla NDM-1 , bla VIM-1 , qnrS1 , or armA , in clinical Enterobacterales, especially in Klebsiella pneumoniae strains from different geographical regions [ 3 – 5 ]. Importantly, the pool of resistance genes carried by IncR replicon plasmids may spread to transferable plasmids through transposition or plasmid recombination events, contributing to the high plasticity of multiple-replicon plasmids [ 2 ].…”

Comparative genomic analyses of IncpA1763‐KPC plasmids

“…The Inc pA1763‐KPC replicon (previously called RepB Rep_3‐family is found in plasmid p628‐KPC [1]) manifests as an iteron‐regulated replicon, for which its iterons (10 or 11 copies of 8‐bp direct repeats) are specifically bound by the replication initiation protein RepA. The Inc pA1763‐KPC replicon often coexists with as an auxiliary replicon with the major replicons (such as IncFII and IncR) to compose multireplicon plasmids, represented by plasmids p0716‐KPC [2] and p721005‐KPC [3], respectively. For instance, p0716‐KPC carries a primary replicon IncFII p0716‐KPC and an auxiliary replicon Inc pA1763‐KPC , and displays a complex chimera backbone containing the IncFII K ‐type maintenance and conjugal transfer regions, an IncFII Y ‐type maintenance region, and the Inc pA1763‐KPC backbone region.…”

“…The Inc pA1763‐KPC replicon as a sole replicon in a plasmid is also capable of guaranteeing the plasmid's stable maintenance for inheritance [3]. Inc pA1763‐KPC plasmids are herein defined as those with a single Inc pA1763‐KPC replicon.…”