| Content Provider | Springer Nature : BioMed Central |
|---|---|
| Author | D’Souza, Roshan Nguyen, Le Phuong Pinto, Naina A. Lee, Hyunsook Vu, Thao Nguyen Kim, Hoyoung Cho, Hyun Soo Yong, Dongeun |
| Abstract | Background A complex cascade of genes, enzymes, and transcription factors regulates AmpC β-lactamase overexpression. We investigated the network of AmpC β-lactamase overexpression in Klebsiella aerogenes and identified the role of AmpG in resistance to β-lactam agents, including cephalosporins and carbapenems. Methods A transposon mutant library was created for carbapenem-resistant K. aerogenes YMC2008-M09-943034 (KE-Y1) to screen for candidates with increased susceptibility to carbapenems, which identified the susceptible mutant derivatives KE-Y3 and KE-Y6. All the strains were subjected to highly contiguous de novo assemblies using PacBio sequencing to investigate the loss of resistance due to transposon insertion. Complementation and knock-out experiments using lambda Red-mediated homologous recombinase and CRISPR–Cas9 were performed to confirm the role of gene of interest. Results In-depth analysis of KE-Y3 and KE-Y6 revealed the insertion of a transposon at six positions in each strain, at which truncation of the AmpG permease gene was common in both. The disruption of the AmpG permease leads to carbapenem susceptibility, which was further confirmed by complementation. We generated an AmpG permease gene knockout using lambda Red-mediated recombineering in K. aerogenes KE-Y1 and a CRISPR–Cas9-mediated gene knockout in multidrug-resistant Klebsiella pneumoniae-YMC/2013/D to confer carbapenem susceptibility. Conclusions These findings suggest that inhibition of the AmpG is a potential strategy to increase the efficacy of β-lactam agents against Klebsiella aerogenes. |
| Related Links | https://ann-clinmicrob.biomedcentral.com/counter/pdf/10.1186/s12941-021-00446-7.pdf |
| Ending Page | 10 |
| Page Count | 10 |
| Starting Page | 1 |
| File Format | HTM / HTML |
| ISSN | 14760711 |
| DOI | 10.1186/s12941-021-00446-7 |
| Journal | Annals of Clinical Microbiology and Antimicrobials |
| Issue Number | 1 |
| Volume Number | 20 |
| Language | English |
| Publisher | BioMed Central |
| Publisher Date | 2021-06-16 |
| Access Restriction | Open |
| Subject Keyword | Medical Microbiology Infectious Diseases Drug Resistance Microbial Genetics and Genomics Biochemistry Parasitology AmpG Carbapenem resistance CRISPR–Cas9 |
| Content Type | Text |
| Resource Type | Article |
| Subject | Microbiology (medical) Infectious Diseases |
| Journal Impact Factor | 4.6/2023 |
| 5-Year Journal Impact Factor | 5.3/2023 |
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