Table 1.
Minimal inhibitory concentrations of carbapenems for K. pneumoniae.
Fig 1.
(A) FDR and log2FC were used to screen for DEGs. (B) KEGG pathway enrichment analysis of DEGs. Fourteen significant enriched pathways (P < 0.05) are represented. (C) GO analysis of DEGs. DEGs are divided into three categories: biological processes, molecular functions, and cellular components. The Y-axis represents the number of genes corresponding to each classification entry and the X-axis represents meaningful enrichment pathways (P < 0.05).
Table 2.
Differentially expressed sRNAs of KPC-2-producing CRKP.
Fig 2.
Validation of differentially expressed sRNAs in KPC-2-producing CRKP.
The expressions of sRNA51 (A), sRNA93 (B), sRNA156 (C), sRNA398 (D), and sRNA418 (E) in CSKP and KPC-2-producing CRKP were detected by qRT-PCR. Data are expressed as mean ± SD (n = 10), * P < 0.05 and *** P < 0.001. (F) The secondary structures and gene sequences of sRNA51.
Table 3.
The ORF values of sRNA51.
Fig 3.
Validation of the target mRNA of sRNA51.
The RNA (A) and protein (B) expression level of acrA in CSKP and KPC-2-producing CRKP were detected by qRT-PCR and western blotting (n = 10). (C) A schematic of the binding of sRNA51 and acrA. The RNA expression levels of sRNA51 (D) and acrA (E) in KPC-2-producing CRKP and KPC-2+sRNA51 strains were detected by qRT-PCR (n = 3). (F) Western blotting analysis of the protein expression level of acrA in KPC-2-producing CRKP and KPC-2+sRNA51 strains. The data presented are representative images of three independent experiments with similar results (S3 Fig). (G) Resistance of KPC-2 producing CRKP and KPC-2+sRNA51 strains to meropenem, ertapenem and imipenem was detected by microbroth dilution. Data are expressed as mean ± SD, * P < 0.05, **P < 0.01 and ***P < 0.001.
Table 4.
The MIC of carbapenems for sRNA51-overexpressing strains of KPC-2-producing CRKP.
Fig 4.
Mechanism of sRNA51 regulation carbapenems resistance of KPC-2-producing CRKP.
qRT-PCR (A) and western blotting (B) detected expression levels of acrA in KPC-2+sRNA51 and sRNA51+acrA strains (n = 3). The data presented are representative images of three independent experiments with similar results (S3 Fig). Detection of resistance to meropenem (C) and ertapenem (D) in KPC-2+sRNA51 and sRNA51+acrA strains by microbroth dilution. Data are represented as the mean ± SD, # P < 0.05 indicates a significant difference compared with KPC-2-producing CRKP and *P < 0.05 indicates a significant difference compared with KPC-2+sRNA51.
Table 5.
The MIC of carbapenems for overexpressed acrA in KPC-2+sRNA51 strains.
Table 6.
Primer sequences used for qRT-PCR.