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      Contributions of the oligopeptide permeases in multistep of Vibrio alginolyticus pathogenesis

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          Abstract

          Vibrio alginolyticus has been associated with several diseases of cultivated marine animals, and has led to considerable economic losses. The oligopeptide permease (Opp) has been proven to play a variety of important roles in nutrition and virulence in several bacteria. In our previous research, the opp gene cluster was identified in Vibrio alginolyticus with transcriptome sequence, which also indicated that the Opp system might play roles in the regulation of adhesion. In this study, the relationship between V. alginolyticus virulence and the opp gene cluster was determined using gene silencing followed by RT‐qPCR, in vitro adhesion assay, growth curves detection in the presence of glutathione (GSH) as a toxic substrate, hemolysis assay, biofilm assay, and artificial infection. Silencing these genes led to deficiencies in adhesion, peptide internalization, biofilm production, hemolytic activity, and virulence. The expression levels of hapr, hapa, tlh, and hlya, which are important genes closely related to the hemolytic activity of Vibrio, were significantly downregulated in all of the RNAi groups. Furthermore, the expression of oppA, oppB, oppC, oppD, and oppF was significantly influenced by temperature, starvation, and pH. These results indicate that (1) oppABCDF contributed in multistep of V. alginolyticus pathogenesis, including adhesion, biofilm production, and hemolytic activity; (2) oppABCDF was sensitive to different temperatures, changes in pH, and increased starvation time.

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          Genetic requirements for mycobacterial survival during infection.

          Despite the importance of tuberculosis as a public health problem, we know relatively little about the molecular mechanisms used by the causative organism, Mycobacterium tuberculosis, to persist in the host. To define these mechanisms, we have mutated virtually every nonessential gene of M. tuberculosis and determined the effect disrupting each gene on the growth rate of this pathogen during infection. A total of 194 genes that are specifically required for mycobacterial growth in vivo were identified. The behavior of these mutants provides a detailed view of the changing environment that the bacterium encounters as infection proceeds. A surprisingly large fraction of these genes are unique to mycobacteria and closely related species, indicating that many of the strategies used by this unusual group of organisms are fundamentally different from other pathogens
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            Bacterial adhesion and entry into host cells.

            Successful establishment of infection by bacterial pathogens requires adhesion to host cells, colonization of tissues, and in certain cases, cellular invasion-followed by intracellular multiplication, dissemination to other tissues, or persistence. Bacteria use monomeric adhesins/invasins or highly sophisticated macromolecular machines such as type III secretion systems and retractile type IV pili to establish a complex host/pathogen molecular crosstalk that leads to subversion of cellular functions and establishment of disease.
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              Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid.

              Construction and characterization of a class of multicopy plasmid cloning vehicles containing the replication system of miniplasmid P15A are described. The constructed plasmids have cleavage sites within antibiotic resistance genes for a variety of commonly employed site-specific endonucleases, permitting convenient use of the insertional inactivation procedure for the selection of clones that contain hybrid DNA molecules. Although the constructed plasmids showed DNA sequence homology with the ColE1 plasmid within the replication region, were amplifiable by chloramphenicol or spectinomycin, required DNA polymerase I for replication, and shared other replication properties with ColE1, they were nevertheless compatible with ColE1. P15A-derived plasmids were not self-transmissible and were mobilized poorly by Hfr strains; however, mobilization was complemented by the presence of a ColE1 plasmid within the same cell.
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                Author and article information

                Contributors
                yanqp@jmu.edu.cn
                Journal
                Microbiologyopen
                Microbiologyopen
                10.1002/(ISSN)2045-8827
                MBO3
                MicrobiologyOpen
                John Wiley and Sons Inc. (Hoboken )
                2045-8827
                17 July 2017
                October 2017
                : 6
                : 5 ( doiID: 10.1002/mbo3.2017.6.issue-5 )
                : e00511
                Affiliations
                [ 1 ] Key Laboratory of Healthy Mariculture for the East China Sea Fisheries College Ministry of Agriculture Jimei University Xiamen China
                [ 2 ] State Key Laboratory of Large Yellow Croaker Breeding Ningde China
                [ 3 ] College of Ocean & Earth Sciences Xiamen University Xiamen China
                Author notes
                [*] [* ] Correspondence

                Qingpi Yan, Fisheries College, Jimei University, Xiamen, China.

                Email: yanqp@ 123456jmu.edu.cn

                [†]

                Co‐first author.

                Author information
                http://orcid.org/0000-0001-8698-3121
                Article
                MBO3511
                10.1002/mbo3.511
                5635161
                28714216
                4d71caa6-fe59-415a-9f1e-3a6401c3d0e8
                © 2017 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

                This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 28 April 2017
                : 25 May 2017
                : 30 May 2017
                Page count
                Figures: 7, Tables: 0, Pages: 9, Words: 6843
                Funding
                Funded by: National Natural Science Foundation of China
                Funded by: Science and Technology Major/Special Project of Fujian Province
                Award ID: 2016NZ0001‐3
                Funded by: Fujian Provincial Department of Science & Technology
                Award ID: JA15289
                Funded by: Natural Science Foundation of Fujian Province
                Award ID: 2016J05080
                Funded by: Scientific Research Fund of Fujian Provincial Department of Education
                Award ID: JA15292
                Funded by: Jimei University
                Award ID: C515044
                Categories
                Original Research
                Original Research
                Custom metadata
                2.0
                mbo3511
                October 2017
                Converter:WILEY_ML3GV2_TO_NLMPMC version:5.2.1 mode:remove_FC converted:11.10.2017

                Microbiology & Virology
                adhesion,biofilm,hemolytic activity,opp system,pathogenesis,rnai,vibrio alginolyticus

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