Open AccessExpression of fluorescent proteins in Lactobacillus rhamnosus to study host–microbe and microbe–microbe interactions
Author(s) -
Spacova Irina,
Lievens Elke,
Verhoeven Tine,
Steenackers Hans,
Vanderleyden Jos,
Lebeer Sarah,
Petrova Mariya I.
Publication year - 2018
Publication title -
microbial biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.287
H-Index - 74
ISSN - 1751-7915
DOI - 10.1111/1751-7915.12872
Subject(s) - lactobacillus rhamnosus , probiotic , microbiology and biotechnology , biology , lactobacillus , biofilm , heterologous , green fluorescent protein , bacteria , biochemistry , gene , genetics
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 mT ag BFP 2 and mC herry 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 mT ag BFP 2 and mC herry 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.