ArticlePloS one2016
In Vivo and In Vitro Detection of Luminescent and Fluorescent Lactobacillus reuteri and Application of Red Fluorescent mCherry for Assessing Plasmid Persistence.
Article in PloS one, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 37 citations in OpenAlex.
- Harnessing endogenous CRISPR-Cas9 for inducible genetic engineering ofApplied and environmental microbiology · 2026Article
- Probiotic Bacteria as Therapeutics and Biohybrid Drug Carriers: Advances, Design Strategies, and Future Outlook.ACS applied bio materials · 2025Review
- Limosilactobacillus reuteri enables oral-to-systemic absorption of iberiotoxin for treatment of collagen-induced arthritis in rats.Microbial cell factories · 2025Article
- B Cells as a Host of Persistent Salmonella Typhimurium.Immunology · 2025Article
- Recent advances in genetic tools for engineering probiotic lactic acid bacteria.Bioscience reports · 2023Article
- RapidBio-protocol · 2022Article
- Introduction of Plasmid to the Murine Gut via Consumption of anRegenerative engineering and translational medicine · 2022Article
- Engineering of Vaginal Lactobacilli to Express Fluorescent Proteins Enables the Analysis of Their Mixture in Nanofibers.International journal of molecular sciences · 2021Article
- A novel universal nano-luciferase-involved reporter system for long-term probing food-borne probiotics and pathogenic bacteria in mice byRSC advances · 2020Article
- Combining Modules for Versatile and Optimal Labeling of Lactic Acid Bacteria: Two pMV158-Family Promiscuous Replicons, a Pneumococcal System for Constitutive or Inducible Gene Expression, and Two Fluorescent Proteins.Frontiers in microbiology · 2019Article
- In vivo bioluminescence imaging of the spatial and temporal colonization of lactobacillus plantarum 423 and enterococcus mundtii ST4SA in the intestinal tract of mice.BMC microbiology · 2018Article
- Construction of a Bioluminescent Labelling Plasmid Vector for Bifidobacteria.Korean journal for food science of animal resources · 2018Article
- In Vitro Transcriptome Response to a Mixture ofInternational journal of molecular sciences · 2018Article
- Tracking of Engineered Bacteria In Vivo Using Nonstandard Amino Acid Incorporation.ACS synthetic biology · 2018Article
- Synthetic Biology Approaches to Engineer Probiotics and Members of the Human Microbiota for Biomedical Applications.Annual review of biomedical engineering · 2018Review
- Expression of fluorescent proteins in Lactobacillus rhamnosus to study host-microbe and microbe-microbe interactions.Microbial biotechnology · 2018Article
- Genetic Tools for the Enhancement of Probiotic Properties.Microbiology spectrum · 2017Review
- Monitoring of BioluminescentKorean journal for food science of animal resources · 2017Article
- Development of a Click Beetle Luciferase Reporter System for Enhanced Bioluminescence Imaging ofFrontiers in microbiology · 2017Article
- Fluorescent reporter systems for tracking probiotic lactic acid bacteria and bifidobacteria.World journal of microbiology & biotechnology · 2016Review
Corrections and comments
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Lactobacillus reuteri is a symbiont that inhabits the gastrointestinal (GI) tract of mammals, and several strains are used as probiotics. After introduction of probiotic strains in a complex ecosystem like the GI tract, keeping track of them is a challenge. The main objectives of this study were to introduce reporter proteins that would enable in vivo and in vitro detection of L. reuteri and increase knowledge about its interactions with the host. We describe for the first time cloning of codon-optimized reporter genes encoding click beetle red luciferase (CBRluc) and red fluorescent protein mCherry in L. reuteri strains ATCC PTA 6475 and R2LC. The plasmid persistence of mCherry-expressing lactobacilli was evaluated by both flow cytometry (FCM) and conventional plate count (PC), and the plasmid loss rates measured by FCM were lower overall than those determined by PC. Neutralization of pH and longer induction duration significantly improved the mCherry signal. The persistency, dose-dependent signal intensity and localization of the recombinant bacteria in the GI tract of mice were studied with an in vivo imaging system (IVIS), which allowed us to detect fluorescence from 6475-CBRluc-mCherry given at a dose of 1×1010 CFU and luminescence signals at doses ranging from 1×105 to 1×1010 CFU. Both 6475-CBRluc-mCherry and R2LC-CBRluc were localized in the colon 1 and 2 h after ingestion, but the majority of the latter were still found in the stomach, possibly reflecting niche specificity for R2LC. Finally, an in vitro experiment showed that mCherry-producing R2LC adhered efficiently to the intra cellular junctions of cultured IPEC-J2 cells. In conclusion, the two reporter genes CBRluc and mCherry were shown to be suitable markers for biophotonic imaging (BPI) of L. reuteri and may provide useful tools for future studies of in vivo and in vitro interactions between the bacteria and the host.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.