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A streamlined method for transposon mutagenesis of Rickettsia parkeri yields numerous mutations that impact infection

View ORCID ProfileRebecca L. Lamason, Natasha M. Kafai, View ORCID ProfileMatthew D. Welch
doi: https://doi.org/10.1101/277160
Rebecca L. Lamason
1Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley,CA
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  • For correspondence: rlamason@mit.edu welch@berkeley.edu
Natasha M. Kafai
1Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley,CA
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Matthew D. Welch
1Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley,CA
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  • For correspondence: rlamason@mit.edu welch@berkeley.edu
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Abstract

The rickettsiae are obligate intracellular alphaproteobacteria that exhibit a complex infectious life cycle in both arthropod and mammalian hosts. As obligate intracellular bacteria, Rickettsia are highly adapted to living inside a variety of host cells, including vascular endothelial cells during mammalian infection. Although it is assumed that the rickettsiae produce numerous virulence factors that usurp or disrupt various host cell pathways, they have been challenging to genetically manipulate to identify the key bacterial factors that contribute to infection. Motivated to overcome this challenge, we sought to expand the repertoire of available rickettsial loss-of-function mutants, using an improved mariner-based transposon mutagenesis scheme. Here, we present the isolation of over 100 transposon mutants in the spotted fever group species Rickettsia parkeri. These mutants targeted genes implicated in a variety of pathways, including bacterial replication and metabolism, hypothetical proteins, the type IV secretion system, as well as factors with previously established roles in host cell interactions and pathogenesis. Given the need to identify critical virulence factors, forward genetic screens such as this will provide an excellent platform to more directly investigate rickettsial biology and pathogenesis.

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Posted March 08, 2018.
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A streamlined method for transposon mutagenesis of Rickettsia parkeri yields numerous mutations that impact infection
Rebecca L. Lamason, Natasha M. Kafai, Matthew D. Welch
bioRxiv 277160; doi: https://doi.org/10.1101/277160
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A streamlined method for transposon mutagenesis of Rickettsia parkeri yields numerous mutations that impact infection
Rebecca L. Lamason, Natasha M. Kafai, Matthew D. Welch
bioRxiv 277160; doi: https://doi.org/10.1101/277160

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