LuxCDE-luxAB-based promoter reporter system to monitor the Yersinia enterocolitica O:3 gene expression in vivo
| Autor: | Ataç Uzel, Elif Bozcal, Melih Dagdeviren, Mikael Skurnik |
|---|---|
| Přispěvatelé: | Department of Bacteriology and Immunology, Medicum, Mikael Skurnik / Principal Investigator, Clinicum, HUSLAB, Ege Üniversitesi |
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Lipopolysaccharides HOST Operon Molecular biology Physiology Yersinia Enterocolitica lcsh:Medicine Artificial Gene Amplification and Extension LIPOPOLYSACCHARIDE Pathology and Laboratory Medicine Polymerase Chain Reaction Biochemistry Mice Genes Reporter Immune Physiology Gene cluster INFECTION Medicine and Health Sciences lcsh:Science Yersinia enterocolitica SUICIDE VECTOR Promoter Regions Genetic Recombination Genetic Mice Inbred BALB C Multidisciplinary biology CONSTRUCTION O Antigens Yersinia 3. Good health Bacterial Pathogens Enzymes Medical Microbiology Multigene Family Female Bioreporter Pathogens Anatomy Oxidoreductases Luciferase Research Article Plasmids Yersinia Infections Virulence Factors 030106 microbiology ANTIGEN Virulence DNA construction Microbiology 03 medical and health sciences OUTER CORE Animals GRAM-NEGATIVE BACTERIA Gene Microbial Pathogens Bacteria lcsh:R SEROTYPE O-3 Organisms Biology and Life Sciences Proteins Promoter Gene Expression Regulation Bacterial biology.organism_classification Research and analysis methods Gastrointestinal Tract Molecular biology techniques Plasmid Construction Mutation Enzymology VIRULENCE lcsh:Q 3111 Biomedicine Digestive System Spleen Genome Bacterial Cloning |
| Zdroj: | PLoS ONE PLoS ONE, Vol 12, Iss 2, p e0172877 (2017) |
| ISSN: | 1932-6203 |
| Popis: | WOS: 000394688200160 PubMed ID: 28235077 It is crucial to understand the in vitro and in vivo regulation of the virulence factor genes of bacterial pathogens. In this study, we describe the construction of a versatile reporter system for Yersinia enterocolitica serotype O:3 (YeO3) based on the luxCDABE operon. In strain YeO3-luxCDE we integrated the luciferase substrate biosynthetic genes, luxCDE, into the genome of the bacterium so that the substrate is constitutively produced. The luxAB genes that encode the luciferase enzyme were cloned into a suicide vector to allow cloning of any promoter-containing fragment upstream the genes. When the obtained suicide-construct is mobilized into YeO3-luxCDE bacteria, it integrates into the recipient genome via homologous recombination between the cloned promoter fragment and the genomic promoter sequence and thereby generates a single-copy and stable promoter reporter. Lipopolysaccharide (LPS) O-antigen (O-ag) and outer core hexasaccharide (OC) of YeO3 are virulence factors necessary to colonization of the intestine and establishment of infection. To monitor the activities of the OC and O-ag gene cluster promoters we constructed the reporter strains YeO3-P-oc Center for International Mobility (CIMO) fellowship [KM -13-8757, TM-14-9081]; FEMS This work was supported by the Center for International Mobility (CIMO) fellowship grants KM -13-8757 and TM-14-9081 (http://www.cimo.fi/ frontpage), FEMS (http://tems-microbiology.org/ fems-work/grants/fems-research-grants/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. |
| Databáze: | OpenAIRE |
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