Defining Gene-Phenotype Relationships in <named-content content-type="genus-species">Acinetobacter baumannii</named-content> through One-Step Chromosomal Gene Inactivation

ABSTRACT Rates of infection with hospital-acquired Acinetobacter baumannii have exploded over the past decade due to our inability to limit persistence and effectively treat disease. A. baumannii quickly acquires antibiotic resistance, and its genome encodes mechanisms to tolerate biocides and desic...

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Autores principales:	Ashley T. Tucker, Emily M. Nowicki, Joseph M. Boll, Gregory A. Knauf, Nora C. Burdis, M. Stephen Trent, Bryan W. Davies
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2014
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/ecc7c7d183c04d46929d9e310e155021
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spelling	oai:doaj.org-article:ecc7c7d183c04d46929d9e310e1550212021-11-15T15:47:22ZDefining Gene-Phenotype Relationships in <named-content content-type="genus-species">Acinetobacter baumannii</named-content> through One-Step Chromosomal Gene Inactivation10.1128/mBio.01313-142150-7511https://doaj.org/article/ecc7c7d183c04d46929d9e310e1550212014-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01313-14https://doaj.org/toc/2150-7511ABSTRACT Rates of infection with hospital-acquired Acinetobacter baumannii have exploded over the past decade due to our inability to limit persistence and effectively treat disease. A. baumannii quickly acquires antibiotic resistance, and its genome encodes mechanisms to tolerate biocides and desiccation, which enhance its persistence in hospital settings. With depleted antibiotic options, new methods to treat A. baumannii infections are desperately needed. A comprehensive understanding detailing A. baumannii cellular factors that contribute to its resiliency at genetic and mechanistic levels is vital to the development of new treatment options. Tools to rapidly dissect the A. baumannii genome will facilitate this goal by quickly advancing our understanding of A. baumannii gene-phenotype relationships. We describe here a recombination-mediated genetic engineering (recombineering) system for targeted genome editing of A. baumannii. We have demonstrated that this system can perform directed mutagenesis on wide-ranging genes and operons and is functional in various strains of A. baumannii, indicating its broad application. We utilized this system to investigate key gene-phenotype relationships in A. baumannii biology important to infection and persistence in hospitals, including oxidative stress protection, biocide resistance mechanisms, and biofilm formation. In addition, we have demonstrated that both the formation and movement of type IV pili play an important role in A. baumannii biofilm. IMPORTANCE Acinetobacter baumannii is the causative agent of hospital-acquired infections, including pneumonia and serious blood and wound infections. A. baumannii is an emerging pathogen and has become a threat to public health because it quickly develops antibiotic resistance, making treatment difficult or impossible. While the threat of A. baumannii is well recognized, our understanding of even its most basic biology lags behind. Analysis of A. baumannii cellular functions to identify potential targets for drug development has stalled due in part to laborious genetic techniques. Here we have pioneered a novel recombineering system that facilitates efficient genome editing in A. baumannii by single PCR products. This technology allows for rapid genome editing to quickly ascertain gene-phenotype relationships. To demonstrate the power of recombineering in dissecting A. baumannii biology, we use this system to establish key gene-phenotype relationships important to infection and persistence in hospitals, including oxidative stress protection, biocide resistance, and biofilm formation.Ashley T. TuckerEmily M. NowickiJoseph M. BollGregory A. KnaufNora C. BurdisM. Stephen TrentBryan W. DaviesAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 5, Iss 4 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Ashley T. Tucker Emily M. Nowicki Joseph M. Boll Gregory A. Knauf Nora C. Burdis M. Stephen Trent Bryan W. Davies Defining Gene-Phenotype Relationships in <named-content content-type="genus-species">Acinetobacter baumannii</named-content> through One-Step Chromosomal Gene Inactivation
description	ABSTRACT Rates of infection with hospital-acquired Acinetobacter baumannii have exploded over the past decade due to our inability to limit persistence and effectively treat disease. A. baumannii quickly acquires antibiotic resistance, and its genome encodes mechanisms to tolerate biocides and desiccation, which enhance its persistence in hospital settings. With depleted antibiotic options, new methods to treat A. baumannii infections are desperately needed. A comprehensive understanding detailing A. baumannii cellular factors that contribute to its resiliency at genetic and mechanistic levels is vital to the development of new treatment options. Tools to rapidly dissect the A. baumannii genome will facilitate this goal by quickly advancing our understanding of A. baumannii gene-phenotype relationships. We describe here a recombination-mediated genetic engineering (recombineering) system for targeted genome editing of A. baumannii. We have demonstrated that this system can perform directed mutagenesis on wide-ranging genes and operons and is functional in various strains of A. baumannii, indicating its broad application. We utilized this system to investigate key gene-phenotype relationships in A. baumannii biology important to infection and persistence in hospitals, including oxidative stress protection, biocide resistance mechanisms, and biofilm formation. In addition, we have demonstrated that both the formation and movement of type IV pili play an important role in A. baumannii biofilm. IMPORTANCE Acinetobacter baumannii is the causative agent of hospital-acquired infections, including pneumonia and serious blood and wound infections. A. baumannii is an emerging pathogen and has become a threat to public health because it quickly develops antibiotic resistance, making treatment difficult or impossible. While the threat of A. baumannii is well recognized, our understanding of even its most basic biology lags behind. Analysis of A. baumannii cellular functions to identify potential targets for drug development has stalled due in part to laborious genetic techniques. Here we have pioneered a novel recombineering system that facilitates efficient genome editing in A. baumannii by single PCR products. This technology allows for rapid genome editing to quickly ascertain gene-phenotype relationships. To demonstrate the power of recombineering in dissecting A. baumannii biology, we use this system to establish key gene-phenotype relationships important to infection and persistence in hospitals, including oxidative stress protection, biocide resistance, and biofilm formation.
format	article
author	Ashley T. Tucker Emily M. Nowicki Joseph M. Boll Gregory A. Knauf Nora C. Burdis M. Stephen Trent Bryan W. Davies
author_facet	Ashley T. Tucker Emily M. Nowicki Joseph M. Boll Gregory A. Knauf Nora C. Burdis M. Stephen Trent Bryan W. Davies
author_sort	Ashley T. Tucker
title	Defining Gene-Phenotype Relationships in <named-content content-type="genus-species">Acinetobacter baumannii</named-content> through One-Step Chromosomal Gene Inactivation
title_short	Defining Gene-Phenotype Relationships in <named-content content-type="genus-species">Acinetobacter baumannii</named-content> through One-Step Chromosomal Gene Inactivation
title_full	Defining Gene-Phenotype Relationships in <named-content content-type="genus-species">Acinetobacter baumannii</named-content> through One-Step Chromosomal Gene Inactivation
title_fullStr	Defining Gene-Phenotype Relationships in <named-content content-type="genus-species">Acinetobacter baumannii</named-content> through One-Step Chromosomal Gene Inactivation
title_full_unstemmed	Defining Gene-Phenotype Relationships in <named-content content-type="genus-species">Acinetobacter baumannii</named-content> through One-Step Chromosomal Gene Inactivation
title_sort	defining gene-phenotype relationships in <named-content content-type="genus-species">acinetobacter baumannii</named-content> through one-step chromosomal gene inactivation
publisher	American Society for Microbiology
publishDate	2014
url	https://doaj.org/article/ecc7c7d183c04d46929d9e310e155021
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Defining Gene-Phenotype Relationships in <named-content content-type="genus-species">Acinetobacter baumannii</named-content> through One-Step Chromosomal Gene Inactivation

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