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      Interplay between OmpA and RpoN Regulates Flagellar Synthesis in Stenotrophomonas maltophilia

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          Abstract

          OmpA, which encodes outer membrane protein A (OmpA), is the most abundant transcript in Stenotrophomonas maltophilia based on transcriptome analyses. The functions of OmpA, including adhesion, biofilm formation, drug resistance, and immune response targets, have been reported in some microorganisms, but few functions are known in S. maltophilia. This study aimed to elucidate the relationship between OmpA and swimming motility in S. maltophilia. KJΔOmpA, an ompA mutant, displayed compromised swimming and failure of conjugation-mediated plasmid transportation. The hierarchical organization of flagella synthesis genes in S. maltophilia was established by referencing the Pseudomonas aeruginosa model and was confirmed using mutant construction, qRT-PCR, and functional assays. Distinct from the P. aeruginosa model, rpoN, rather than fleQ and fliA, was at the top of the flagellar regulatory cascade in S. maltophilia. To elucidate the underlying mechanism responsible for ΔompA-mediated swimming compromise, transcriptome analysis of KJ and KJΔOmpA was performed and revealed rpoN downregulation in KJΔOmpA as the key element. The involvement of rpoN in ΔompA-mediated swimming compromise was verified using rpoN complementation, qRT-PCR, and function assays. Collectively, OmpA, which contributes to bacterial conjugation and swimming, is a promising target for adjuvant design in S. maltophilia.

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          Most cited references61

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          Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

          The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data. Copyright 2001 Elsevier Science (USA).
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            The bacterial cell envelope.

            The bacteria cell envelope is a complex multilayered structure that serves to protect these organisms from their unpredictable and often hostile environment. The cell envelopes of most bacteria fall into one of two major groups. Gram-negative bacteria are surrounded by a thin peptidoglycan cell wall, which itself is surrounded by an outer membrane containing lipopolysaccharide. Gram-positive bacteria lack an outer membrane but are surrounded by layers of peptidoglycan many times thicker than is found in the gram-negatives. Threading through these layers of peptidoglycan are long anionic polymers, called teichoic acids. The composition and organization of these envelope layers and recent insights into the mechanisms of cell envelope assembly are discussed.
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              Molecular Basis of Bacterial Outer Membrane Permeability Revisited

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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Microorganisms
                Microorganisms
                microorganisms
                Microorganisms
                MDPI
                2076-2607
                04 June 2021
                June 2021
                : 9
                : 6
                : 1216
                Affiliations
                [1 ]Division of Infectious Disease, Far Eastern Memorial Hospital, New Taipei City 220, Taiwan; liaochunhsing@ 123456gmail.com
                [2 ]Department of Medicine, National Yang Ming Chiao Tung University, Taipei 112, Taiwan; ytlin8@ 123456vghtpe.gov.tw
                [3 ]Department of Biotechnology and Laboratory Science in Medicine, National Yang Ming Chiao Tung University, Taipei 112, Taiwan; kartd0087603@ 123456gmail.com (C.-L.C.); toe3273917@ 123456outlook.com (H.-H.H.)
                [4 ]Division of Infectious Diseases, Department of Medicine, Taipei Veterans General Hospital, Taipei 112, Taiwan
                [5 ]Department of Pathology and Laboratory Medicine, Taipei Veterans General Hosiptal, Taipei 112, Taiwan; lilh@ 123456vghtpe.gov.tw
                [6 ]Ph.D. Program in Medical Biotechnology, Taipei Medical University, Taipei 110, Taiwan
                Author notes
                [* ]Correspondence: tcyang@ 123456nycu.edu.tw
                [†]

                Liao, C.-H. and Chang, C.-L. contributed equally to this work.

                Article
                microorganisms-09-01216
                10.3390/microorganisms9061216
                8229486
                34199787
                b20f292a-2ac1-49d1-9863-fefbe5257412
                © 2021 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( https://creativecommons.org/licenses/by/4.0/).

                History
                : 21 April 2021
                : 02 June 2021
                Categories
                Article

                stenotrophomonas maltophilia,ompa,swimming,flagellum
                stenotrophomonas maltophilia, ompa, swimming, flagellum

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