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Transposon mutagenesis screen in Klebsiella pneumoniae identifies genetic determinants required for growth in human urine and serum

Jessica Gray, View ORCID ProfileVon Vergel L Torres, View ORCID ProfileEmily CA Goodall, Karl A Dunne, View ORCID ProfileChris Icke, View ORCID ProfileJessica L Rooke, View ORCID ProfileThamarai Schneiders, View ORCID ProfileAdam F Cunningham, Jeffrey A Cole, View ORCID ProfileIan R Henderson
doi: https://doi.org/10.1101/2023.05.31.543172
Jessica Gray
1Institute for Molecular Bioscience, University of Queensland, St. Lucia, Queensland, Australia
2Institute of Microbiology and Infection, University of Birmingham, Birmingham, United Kingdom
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Von Vergel L Torres
1Institute for Molecular Bioscience, University of Queensland, St. Lucia, Queensland, Australia
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Emily CA Goodall
1Institute for Molecular Bioscience, University of Queensland, St. Lucia, Queensland, Australia
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Karl A Dunne
2Institute of Microbiology and Infection, University of Birmingham, Birmingham, United Kingdom
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Chris Icke
1Institute for Molecular Bioscience, University of Queensland, St. Lucia, Queensland, Australia
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Jessica L Rooke
1Institute for Molecular Bioscience, University of Queensland, St. Lucia, Queensland, Australia
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Thamarai Schneiders
3Division of Infection Medicine, University of Edinburgh, Edinburgh, United Kingdom
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Adam F Cunningham
4Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, United Kingdom
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  • ORCID record for Adam F Cunningham
Jeffrey A Cole
2Institute of Microbiology and Infection, University of Birmingham, Birmingham, United Kingdom
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Ian R Henderson
1Institute for Molecular Bioscience, University of Queensland, St. Lucia, Queensland, Australia
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  • ORCID record for Ian R Henderson
  • For correspondence: [email protected]
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Abstract

Klebsiella pneumoniae is a global public health concern due to the rising myriad of hypervirulent and multi-drug resistant clones both alarmingly associated with high mortality. The molecular microbial genetics underpinning these recalcitrant K. pneumoniae infections is unclear, coupled with the emergence of lineages resistant to nearly all present day clinically important antimicrobials. In this study, we performed a genome-wide screen in K. pneumoniae ECL8, a member of the endemic K2-ST375 pathotype most often reported in Asia, to define genes essential for growth in a nutrient-rich laboratory medium (Luria-Bertani medium), human urine and serum. Through transposon directed insertion-site sequencing (TraDIS), a total of 427 genes were identified as essential for growth on LB agar, whereas transposon insertions in 11 and 144 genes decreased fitness for growth in either urine or serum, respectively. Genome-wide functional studies like these provide further knowledge on the genetics of this pathogen but also provide a strong impetus for discovering new antimicrobial targets to improve current therapeutic options for K. pneumoniae infections.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • https://drive.google.com/drive/folders/1xTuNvlcuN1Vztn5_WteEuc9zsgT9CZS1?usp=sharing

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted June 01, 2023.
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Transposon mutagenesis screen in Klebsiella pneumoniae identifies genetic determinants required for growth in human urine and serum
Jessica Gray, Von Vergel L Torres, Emily CA Goodall, Karl A Dunne, Chris Icke, Jessica L Rooke, Thamarai Schneiders, Adam F Cunningham, Jeffrey A Cole, Ian R Henderson
bioRxiv 2023.05.31.543172; doi: https://doi.org/10.1101/2023.05.31.543172
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Transposon mutagenesis screen in Klebsiella pneumoniae identifies genetic determinants required for growth in human urine and serum
Jessica Gray, Von Vergel L Torres, Emily CA Goodall, Karl A Dunne, Chris Icke, Jessica L Rooke, Thamarai Schneiders, Adam F Cunningham, Jeffrey A Cole, Ian R Henderson
bioRxiv 2023.05.31.543172; doi: https://doi.org/10.1101/2023.05.31.543172

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