Expression of fluorescent proteins in Lactobacillus rhamnosus to study host–microbe and microbe–microbe interactions. Issue 2 (13th October 2017)

• Record Type: Journal Article
• Authors: Spacova, Irina; Lievens, Elke; Verhoeven, Tine; Steenackers, Hans; Vanderleyden, Jos; Lebeer, Sarah; Petrova, Mariya I.
• Is Part Of: Microbial biotechnology. Volume 11:Issue 2(2018:Mar.)
• Page Start: 317
• Page End: 331
• Publication Date: 2017-10-13
• Subjects: probiotics -- fluorescence -- mTagBFP2 -- mCherry -- biofilms -- adhesion; Microbial biotechnology -- Periodicals; Biotechnology; Microbiology; 660.62
• DOI: 10.1111/1751-7915.12872
• Languages: English
• ISSNs: 1751-7915

Abstract

Summary: Probiotic Lactobacillus strains are widely used to benefit human and animal health, although the exact mechanisms behind their interactions with the host and the microbiota are largely unknown. Fluorescent tagging of live probiotic cells is an important tool to unravel their modes of action. In this study, the implementation of different heterologously expressed fluorescent proteins for the labelling of the model probiotic strains Lactobacillus rhamnosus GG (gastrointestinal) and Lactobacillus rhamnosus GR‐1 (vaginal) was explored. Heterologous expression of mTagBFP2 and mCherry resulted in long‐lasting fluorescence of L. rhamnosus GG and GR‐1 cells, using the nisin‐controlled expression (NICE) system. These novel fluorescent strains were then used to study in vitro aspects of their microbe–microbe and microbe–host interactions. Lactobacillus rhamnosus GG and L. rhamnosus GR‐1 expressing mTagBFP2 and mCherry could be visualized in mixed‐species biofilms, where they inhibited biofilm formation by Salmonella Typhimurium– gfpmut3 expressing the green fluorescent protein. Likewise, fluorescent L. rhamnosus GG and L. rhamnosus GR‐1 were implemented for the visualization of their adhesion patterns to intestinal epithelial cell cultures. The fluorescent L. rhamnosus strains developed in this study can therefore serve as novel tools for the study of probiotic interactions with their environment. Abstract : Fluorescent tagging of Lactobacillus probiotic strains is an important tool to unravel their modes of action. In this study, the implementation of heterologously expressed mTagBFP2 and mCherry for the labeling of Lactobacillus rhamnosus GG and Lactobacillus rhamnosus GR‐1 was explored. The novel fluorescent strains were used to study in vitro aspects of probiotoc microbe‐microbe and microbe‐host interactions. The fluorescent L. rhamnosus strains developed in this study can serve as novel tools for the study of probiotic interactions with their environment.