kpc plasmid f33  (INCF)

Journal: eBioMedicine

Article Title: Global emergence of Carbapenem-resistant Hypervirulent Klebsiella pneumoniae driven by an IncFII K34 KPC-2 plasmid

doi: 10.1016/j.ebiom.2025.105627

Figure Lengend Snippet: Genomic features of four KL1-ST23 carbapenem-resistant hypervirulent Klebsiella pneumoniae .

Article Snippet: Among the 174 KPC-encoding genomes out of the 414 global CR-hvKp genomes, IncFII K34 KPC plasmid (24%, 42/174) was the most prevalent IncF KPC plasmid, followed by F33 (9%, 16/174) and F35 (8%, 14/174) plasmids ( g).

Techniques: Plasmid Preparation, CRISPR

Journal: eBioMedicine

Article Title: Global emergence of Carbapenem-resistant Hypervirulent Klebsiella pneumoniae driven by an IncFII K34 KPC-2 plasmid

doi: 10.1016/j.ebiom.2025.105627

Figure Lengend Snippet: Analysis of IncFII K1-34 alleles and global epidemiology of CR-hvKp . a . The classification tree for different IncFII K1-34 alleles was constructed using the Neighbour-Joining method in MEGA and was rooted at the midpoint. Sequences from the copA region of the IncFII K replicon were used in the tree construction. b . The alignment of the IncFII K34 allele with 33 known IncFII K alleles is shown. Dots indicate nucleotides identical to the consensus sequence, and dashes in grey represent gaps. c . The proportion of plasmids harbouring carbapenemase-encoding genes in each IncFII K plasmid type was calculated using data from PLSDB v. 2021_06_23_v2. d . The global distribution of 2,168 hvKp and CR-hvKp genomes with country information available from the NCBI GenBank is illustrated. Countries with more than 20 reported hvKp and CR-hvKp genomes were presented, and the predominant carbapenemase type within the country is highlighted on the top of the bar. e . The percentages of ST23, ST86, and ST65 among 2,343 hvKp and CR-hvKp strains are presented. f. The percentages of carbapenemases found among 414 CR-hvKp strains are detailed. g. The distribution of different IncF plasmid replicons among 174 KPC-encoding CR-hvKp strains is depicted.

Article Snippet: Among the 174 KPC-encoding genomes out of the 414 global CR-hvKp genomes, IncFII K34 KPC plasmid (24%, 42/174) was the most prevalent IncF KPC plasmid, followed by F33 (9%, 16/174) and F35 (8%, 14/174) plasmids ( g).

Techniques: Construct, Sequencing, Plasmid Preparation

Journal: eBioMedicine

Article Title: Global emergence of Carbapenem-resistant Hypervirulent Klebsiella pneumoniae driven by an IncFII K34 KPC-2 plasmid

doi: 10.1016/j.ebiom.2025.105627

Figure Lengend Snippet: Comparative analysis of KPC plasmids in CR-hvKp clinical isolates and phylogenetic analysis of ST23 hvKp and CR-hvKp strains . a . Circular comparison of the three IncFII K34 plasmids (pKPC-H39, pKPC-DD02357 and pKPC-DD01665) in CR-hvKp with pDD02172-2 (CP087613), p1140-KPC (CP047689) and pKP18069-KPC (CP059890). b . Genetic elements of bla KPC-2 on IncFII K34 plasmid. c . Circular comparison of the IncN plasmid (pKPC-DD02201) in CR-hvKp with pNB5_KPC-2 (CP092655), pSZN_KPC (MH917123) and pECN580 (KF914891). d . Genetic elements of bla KPC-2 on IncN plasmid. Open reading frames (ORFs) are indicated by arrows and coloured based on predicted gene function. The replication-associated genes are denoted as blue arrows. Resistance genes are indicated by red arrows, while insertion sequences are shown by yellow arrows. Genes in the conjugation module are shown by green arrows. Other genes are indicated by grey arrows. e . Time-based phylogenetic analysis of 342 ST23 hvKp and CR-hvKp strains. Colours in columns illustrate region of origin, BAPS cluster, host, source of isolation, K locus type, ICE type, carbapenemase type and presence of replicons. Selected divergence time and 95% CIs are shown at the nodes. All the representative genomes are complete, and strains with red dots on the graph are hvKp and with blue dots are CR-hvKp. The nine CR-hvKp strains harbouring the IncFII K34 bla KPC-2 plasmids are indicated by black arrows.

Article Snippet: Among the 174 KPC-encoding genomes out of the 414 global CR-hvKp genomes, IncFII K34 KPC plasmid (24%, 42/174) was the most prevalent IncF KPC plasmid, followed by F33 (9%, 16/174) and F35 (8%, 14/174) plasmids ( g).

Techniques: Comparison, Plasmid Preparation, Conjugation Assay, Isolation

Journal: eBioMedicine

Article Title: Global emergence of Carbapenem-resistant Hypervirulent Klebsiella pneumoniae driven by an IncFII K34 KPC-2 plasmid

doi: 10.1016/j.ebiom.2025.105627

Figure Lengend Snippet: Conjugation assays of bla KPC-2 bearing plasmids and RNA-seq analysis of transconjugants . Conjugation frequencies of KPC-2 plasmids a from CR-hvKp or CRKp isolates to E. coli J53, and b from E. coli J53 to K. pneumoniae NTUH-K2044. c . Conjugation frequencies of virulence plasmids from K. pneumoniae NTUH-K2044 to E. coli J53. Three biologic repeats were performed for each strain. Conjugation frequencies are calculated by dividing the number of transconjugants (CFU/mL) by the number of recipients (CFU/mL) and indicated by mean ± SD. P values were calculated from student t-tests, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. d . The schematic diagram of the conjugation experiments. The numbers on the arrows are conjugation frequencies. The drugs on the bottom were used for selection in each conjugation experiment. MEM is meropenem, NaN 3 is sodium azide and K 2 TeO 3 is potassium tellurite. Only plasmids were shown in the figure. Plasmid profiles were from the results of Nanopore sequencing. The plasmid profile of HS11286 were from the GenBank. e. Heatmap of the RNA-seq analysis of NTUH-K2044 conjugated with IncFII K34 and IncFII K2 KPC-2 plasmids. Three biologic repeats were performed for each strain. f. Volcano plot of DEGs between NTUH-K2044 conjugated with IncFII K34 and IncFII K2 plasmids. g . Heatmap of expression of conjugation-associated genes.

Article Snippet: Among the 174 KPC-encoding genomes out of the 414 global CR-hvKp genomes, IncFII K34 KPC plasmid (24%, 42/174) was the most prevalent IncF KPC plasmid, followed by F33 (9%, 16/174) and F35 (8%, 14/174) plasmids ( g).

Techniques: Conjugation Assay, RNA Sequencing, Selection, Plasmid Preparation, Nanopore Sequencing, Expressing

Journal: eBioMedicine

Article Title: Global emergence of Carbapenem-resistant Hypervirulent Klebsiella pneumoniae driven by an IncFII K34 KPC-2 plasmid

doi: 10.1016/j.ebiom.2025.105627

Figure Lengend Snippet: The competition experiments of NTUH-K2044 conjugated with IncFII K34 and IncFII K2 bla KPC-2 bearing plasmids . Growth curves of isolates cultured with no meropenem a , 0.5 μg/mL meropenem b , 1 μg/mL meropenem c and 2 μg/mL meropenem d . e . The competition between isolates under conditions with and without 1 μg/mL meropenem. NTUH-K2044::pKPC-H39 and NTUH-K2044::pKPC-HS11286 were used in the competition assays. The copies of bla KPC-2 f and pKPC-H39, pKPC-DD02357 and pKPC-HS11286 in NTUH-K2044 g , respectively. Transcriptional levels of bla KPC-2 in qRT-PCR h and RNA-seq i . The transcriptional levels of bla KPC-2 in qRT-PCR were normalized to the endogenous reference gene rrsE . The normalized read counts of bla KPC-2 in RNA-seq were from the DESeq analysis results. The MICs of NTUH-K2044::pKPC-H39 and NTUH-K2044:: pKPC-DD02357 harbouring IncFII K34 KPC-2 plasmids to meropenem were 8 μg/mL, while NTUH-K2044::pKPC-HS11286 with the IncFII K2 KPC-2 plasmid were 4 μg/mL. Three biologic repeats were performed for each strain. P values were calculated from student t-tests, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Among the 174 KPC-encoding genomes out of the 414 global CR-hvKp genomes, IncFII K34 KPC plasmid (24%, 42/174) was the most prevalent IncF KPC plasmid, followed by F33 (9%, 16/174) and F35 (8%, 14/174) plasmids ( g).

Techniques: Cell Culture, Quantitative RT-PCR, RNA Sequencing, Plasmid Preparation