Improved mini-Tn 7 Delivery Plasmids for Fluorescent Labeling of Stenotrophomonas maltophilia

• DOI: 10.1128/aem.00317-23
• Publication Date: 2023-05-17T14:01:04Z
• Authors: Uwe Mamat; Manuel Hein; Dörte Grella; Claire S. Taylor; Thomas Scholzen; Ifey Alio; Wolfgang R. Streit; Pol Huedo; Xavier Coves; Oscar Conchillo-Solé; Andromeda-Celeste...; Isidre Gibert; Daniel Yero; Ulrich E. Schaible
• Keywords: Stenotrophomonas maltophilia

Fluorescently labeled bacterial cells have become indispensable for many aspects of microbiological research, including studies on biofilm formation as an important virulence factor various opportunistic bacteria environmental origin such Stenotrophomonas maltophilia. Using a Tn7-based genomic integration system, we report the construction improved mini-Tn7 delivery plasmids labeling S. maltophilia with sfGFP, mCherry, tdTomato and mKate2 by expressing their codon-optimized genes from strong, constitutive promoter optimized ribosomal binding site. Transposition transposons into single neutral sites located average 25 nucleotides downstream 3'-end conserved glmS gene different wild-type strains did not any adverse effects fitness fluorescently derivatives. This was demonstrated comparative analyses growth, resistance profiles against 18 antibiotics classes, ability to form biofilms abiotic biotic surfaces, also independent fluorescent protein expressed, in Galleria mellonella. It is shown that elements remained stably integrated genome over prolonged period time absence antibiotic selection pressure. Overall, provide evidence new are valuable tools generating indistinguishable properties parental strains. IMPORTANCE The bacterium nosocomial pathogen can cause bacteremia pneumonia immunocompromised patients high rate mortality. now considered clinically relevant notorious cystic fibrosis but has been isolated lung specimen healthy donors. intrinsic wide range complicates treatment most likely contributes increasing incidence infections worldwide. One virulence-related trait surface, which may result development increased transient phenotypic antimicrobials. significance our work mini-Tn7-based system study mechanisms or host-pathogen interactions live under non-destructive conditions.