Article Figures & Data
Figures
- FIG 1
Detailed genetic features of a virulence-encoding region in the chromosome of strain 16HN-263. (a) Sequence alignment of four complete K. pneumoniae chromosomal sequences, with the chromosome of 16HN-263 being used as a reference. Alignment with the chromosomal sequence of strain HS11286 (GenBank accession no. CP003200) and those of our previously reported strains, CR-HvKP4 and FJ8, indicates that a 100-kbp segment was inserted into the chromosome of strain 16HN-263. (b) Alignment of the 107,648-bp region in the chromosome of strain 16HN-263 with the plasmids p17-16-vir (GenBank accession no. MK191024.1), pCR-HvKP4_VIR (GenBank accession no. CP040540.1), and pLVPK (GenBank accession no. AY378100.1).
- FIG 2
Formation of a circular intermediate by circularization of the insertion segment in the chromosome of strain 16HN-263. (a) Genetic structure of the transposon and its circular intermediate. IS26 elements are shown as open boxes. Green arrows indicate the position and orientation of tnp26. Orange arrows refer to the left and right terminal inverted repeats (IRL and IRR) of the IS26 element. Flags indicate the position and direction of the identified target site duplication (TSD). (b) Genetic structure of the chromosomal fragment after excision of the transposon. (c) Gel electrophoresis of PCR amplicons corresponding to the circular intermediate generated using the reverse primers IS26-F and IS26-R. S depicts the PCR product. (d) Gel electrophoresis of PCR amplicons corresponding to the excision of Tn7074 using the primers Up-F and Up-R. S depicts the PCR product. (e) S1-PFGE profile of strain 16HN-263. S indicates strain 16HN-263.
- FIG 3
Virulence level of different bacterial strains as depicted in a wax worm (Galleria mellonella) larva infection model. (a and b) Survival of wax worm (n = 10) infected by 1 × 106 CFU/ml (a) and 1 × 105 CFU/ml (b) of each K. pneumoniae strain at 48 h. The test strains included K. pneumoniae strain 16HN-263, ST11 CR-HvKP strain CR-HvKP4 (hypervirulence control), ST23 K1 strain HvKP1088 (hypervirulent K. pneumoniae control), and the classic CRKP strain FJ8 (low-virulence control). A log rank (Mantel-Cox) test was performed for the indicated curves. A significant difference (P = 0.0027 [a] and P = 0.0494 [b]) was observed between 16HN-263 and FJ8, while no significant difference was observed between 16HN-263 and CR-HvKP4 or HvKP1088 (P > 0.9999 and P = 0.5417 between 16HN-263 and CR-HvKP4 in panels a and b, respectively; P = 0.3173 and P = 0.5417 between 16HN-263 and HvKP1088 in panels a and b, respectively).
Tables
- TABLE 1
MIC profiles of strain 16HN-263
↵a ATM, aztreonam; CTX, cefotaxime; CAZ, ceftazidime; CMZ, cefmetazole; FEP, cefepime; IMP, imipenem; MEM, meropenem; EM, ertapenem; AMK, amikacin; CIP, ciprofloxacin; AZI, azithromycin; PB, polymyxin B; TGC, tigecycline. All tests were performed in duplicate, and each test included three biological replicates.
FIG S1
Plasmidome analysis of strain 16HN-263 compared to CR-HvKP strains. (A) Alignment of plasmid p16HN-263_KPC with the plasmids pCR-HvKP1-KPC, pCR-HvKP4-KPC, pCR-HvKP5-KPC, and p69-2. (B) Alignment of plasmid p16HN-263_2 with the plasmids pCR-HvKP1-p4, pCR-HvKP4-p4, and pCR-HvKP5-p4. (C) Alignment of plasmid p16HN-263_3 with the plasmids pCR-HvKP1-p5, pCR-HvKP4-p5, and pCR-HvKP5-p5. Download FIG S1, TIF file, 2.0 MB.
Copyright © 2020 Yang et al.
This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.
FIG S2
Confirmation of the integration of a 100-kbp virulence and resistance encoding region into the chromosome. (A) Alignment of nanopore reads (SRR12072214) to chromosome assembly of 16HN-263 (GenBank accession no. CP045263.1). The alignment was generated by BWA-MEM and viewed by IGV. (B) Gel electrophoresis of PCR amplicons across both ends of the integration sites. “Up” depicts the PCR product of the upstream integration site; “Down” depicts the PCR product of the downstream integration site. Download FIG S2, TIF file, 0.2 MB.
Copyright © 2020 Yang et al.
This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.
