Protocol for analyzing the function of the type VI secretion system of the oral symbiont Aggregatibacter aphrophilus in targeting pathobionts.

Autor:	Oscarsson J; Oral Microbiology, Department of Odontology, Umeå University, Umeå, Sweden. Electronic address: jan.oscarsson@umu.se., Bao K; Division of Oral Health and Periodontology, Department of Dental Medicine, Karolinska Institutet, Alfred Nobels Allé 8, 14104 Huddinge, Stockholm, Sweden., Shiratsuchi A; Department of Liberal Arts and Sciences, Graduate School of Medicine, Sapporo Medical University, Sapporo, Hokkaido 060-8556, Japan., Grossmann J; Functional Genomics Center Zurich, ETH Zürich and University of Zürich, Zürich, Switzerland; Swiss Institute of Bioinformatics (SIB) Quartier Sorge-Batiment Amphipole, 1015 Lausanne, Switzerland., Wolski W; Functional Genomics Center Zurich, ETH Zürich and University of Zürich, Zürich, Switzerland; Swiss Institute of Bioinformatics (SIB) Quartier Sorge-Batiment Amphipole, 1015 Lausanne, Switzerland., Aung KM; Department of Molecular Biology and the Umeå Centre for Microbial Research (UCMR), Umeå, Sweden; Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, 90187 Umeå, Sweden., Lindholm M; Oral Microbiology, Department of Odontology, Umeå University, Umeå, Sweden; Division of Oral Health and Periodontology, Department of Dental Medicine, Karolinska Institutet, Alfred Nobels Allé 8, 14104 Huddinge, Stockholm, Sweden., Johansson A; Oral Microbiology, Department of Odontology, Umeå University, Umeå, Sweden., Mowsumi FR; Oral Microbiology, Department of Odontology, Umeå University, Umeå, Sweden., Wai SN; Department of Molecular Biology and the Umeå Centre for Microbial Research (UCMR), Umeå, Sweden; Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, 90187 Umeå, Sweden., Belibasakis GN; Division of Oral Health and Periodontology, Department of Dental Medicine, Karolinska Institutet, Alfred Nobels Allé 8, 14104 Huddinge, Stockholm, Sweden., Bostanci N; Division of Oral Health and Periodontology, Department of Dental Medicine, Karolinska Institutet, Alfred Nobels Allé 8, 14104 Huddinge, Stockholm, Sweden. Electronic address: nagihan.bostanci@ki.se.
Jazyk:	angličtina
Zdroj:	STAR protocols [STAR Protoc] 2024 Dec 20; Vol. 5 (4), pp. 103415. Date of Electronic Publication: 2024 Oct 25.
DOI:	10.1016/j.xpro.2024.103415
Abstrakt:	Here, we present a protocol for evaluating type VI secretion system (T6SS)-dependent fitness of the oral symbiont A. aphrophilus using biofilm competition assays and metaproteomics. We describe steps for designing T6SS-specific mutants. We then detail procedures for using them in competition assays with the pathobiont A. actinomycetemcomitans and in biofilm models, analyzing metaproteomes to assess the impact of the T6SS on multiple pathobionts. The biofilm model is designed to mimic the oral plaque ecosystem and includes seven species. For complete details on the use and execution of this protocol, please refer to Oscarsson et al. 1 . Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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