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ABSTRACT<\/h2>\n\n\n\n
Fluorescently labeled bacterial cells have become indispensable for many aspects of microbiological research, including studies on biofilm formation as an important virulence factor of various opportunistic bacteria of environmental origin such as\u00a0Stenotrophomonas maltophilia<\/em>. Using a Tn7<\/em>-based genomic integration system, we report the construction of improved mini-Tn7<\/em>\u00a0delivery plasmids for labeling of\u00a0S. maltophilia<\/em>\u00a0with sfGFP, mCherry, tdTomato and mKate2 by expressing their codon-optimized genes from a strong, constitutive promoter and an optimized ribosomal binding site. Transposition of the mini-Tn7<\/em>\u00a0transposons into single neutral sites located on average 25 nucleotides downstream of the 3\u2032-end of the conserved\u00a0glmS<\/em>\u00a0gene of different\u00a0S. maltophilia<\/em>\u00a0wild-type strains did not have any adverse effects on the fitness of their fluorescently labeled derivatives. This was demonstrated by comparative analyses of growth, resistance profiles against 18 antibiotics of different classes, the ability to form biofilms on abiotic and biotic surfaces, also independent of the fluorescent protein expressed, and virulence in\u00a0Galleria mellonella<\/em>. It is also shown that the mini-Tn7<\/em>\u00a0elements remained stably integrated in the genome of\u00a0S. maltophilia<\/em>\u00a0over a prolonged period of time in the absence of antibiotic selection pressure. Overall, we provide evidence that the new improved mini-Tn7<\/em>\u00a0delivery plasmids are valuable tools for generating fluorescently labeled\u00a0S. maltophilia<\/em>\u00a0strains that are indistinguishable in their properties from their parental wild-type strains.<\/p>\n\n\n\n I<\/strong>MPORTANCE\u00a0The bacterium\u00a0S. maltophilia\u00a0is an important opportunistic nosocomial pathogen that can cause bacteremia and pneumonia in immunocompromised patients with a high rate of mortality. It is now considered as a clinically relevant and notorious pathogen in cystic fibrosis patients but has also been isolated from lung specimen of healthy donors. The high intrinsic resistance to a wide range of antibiotics complicates treatment and most likely contributes to the increasing incidence of\u00a0S. maltophilia\u00a0infections worldwide. One important virulence-related trait of\u00a0S. maltophilia\u00a0is the ability to form biofilms on any surface, which may result in the development of increased transient phenotypic resistance to antimicrobials. The significance of our work is to provide a mini-Tn7-based labeling system for\u00a0S. maltophilia\u00a0to study the mechanisms of biofilm formation or host-pathogen interactions with live bacteria under non-destructive conditions.<\/p>\n<\/div><\/div>\n\n\n\n <\/p>\n<\/div><\/div>\n\n\n\n <\/p>\n<\/div><\/div>\n","protected":false},"excerpt":{"rendered":" Cellular Microbiology, Priority Research Area Infections, Research Center Borstel, Leibniz Lung Center & Institut de Biotecnologia i de Biomedicina (IBB), Universitat Aut\u00f2noma de Barcelona (UAB) Appl Environ Microbiol. 2023 May 17:e0031723. doi: 10.1128\/aem.00317-23.\u00a0PMID:\u00a037195181.\u00a0 ABSTRACT Fluorescently labeled bacterial cells have become indispensable for many aspects of microbiological research, including studies on biofilm formation as an important […]<\/p>\n","protected":false},"author":65,"featured_media":5175,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[4],"tags":[],"_links":{"self":[{"href":"https:\/\/ibb.uab.cat\/index.php\/wp-json\/wp\/v2\/posts\/5171"}],"collection":[{"href":"https:\/\/ibb.uab.cat\/index.php\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/ibb.uab.cat\/index.php\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/ibb.uab.cat\/index.php\/wp-json\/wp\/v2\/users\/65"}],"replies":[{"embeddable":true,"href":"https:\/\/ibb.uab.cat\/index.php\/wp-json\/wp\/v2\/comments?post=5171"}],"version-history":[{"count":2,"href":"https:\/\/ibb.uab.cat\/index.php\/wp-json\/wp\/v2\/posts\/5171\/revisions"}],"predecessor-version":[{"id":5178,"href":"https:\/\/ibb.uab.cat\/index.php\/wp-json\/wp\/v2\/posts\/5171\/revisions\/5178"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/ibb.uab.cat\/index.php\/wp-json\/wp\/v2\/media\/5175"}],"wp:attachment":[{"href":"https:\/\/ibb.uab.cat\/index.php\/wp-json\/wp\/v2\/media?parent=5171"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/ibb.uab.cat\/index.php\/wp-json\/wp\/v2\/categories?post=5171"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/ibb.uab.cat\/index.php\/wp-json\/wp\/v2\/tags?post=5171"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}![\"\"](\"https:\/\/ibb.uab.cat\/wp-content\/uploads\/Biofilm-formation-Smaltophilia-on-Calu-3-cells-933x1024.jpg\")
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