title: Use of the mCherry Fluorescent Protein To Study Intestinal Colonization by Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 in Mice.
abstract: Lactic acid bacteria (LAB) are natural inhabitants of the gastrointestinal tract (GIT) of humans and animals, and some LAB species receive considerable attention due to their health benefits. Although many papers have been published on probiotic LAB, only a few reports have been published on the migration and colonization of the cells in the GIT. This is due mostly to the lack of efficient reporter systems. In this study, we report on the application of the fluorescent mCherry protein in the in vivo tagging of the probiotic strains Enterococcus mundtii ST4SA and Lactobacillus plantarum 423. The mCherry gene, encoding a red fluorescent protein (RFP), was integrated into a nonfunctional region on the genome of L. plantarum 423 by homologous recombination. In the case of E. mundtii ST4SA, the mCherry gene was cloned into the pGKV223D LAB/Escherichia coli expression vector. Expression of the mCherry gene did not alter the growth rate of the two strains and had no effect on bacteriocin production. Both strains colonized the cecum and colon of mice.
authors.affiliation: Department of Microbiology, University of Stellenbosch, Stellenbosch, South Africa.
authors.forename: Winschau F
authors.initials: WF
authors.lastname: van Zyl
authors.affiliation: Department of Microbiology, University of Stellenbosch, Stellenbosch, South Africa.
authors.forename: Shelly M
authors.initials: SM
authors.lastname: Deane
authors.affiliation: Department of Microbiology, University of Stellenbosch, Stellenbosch, South Africa lmtd@sun.ac.za.
authors.forename: Leon M T
authors.initials: LM
authors.lastname: Dicks
mesh_terms: D000818:Animals; D016983:Enterococcus / Q000737:chemistry / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D005260:Female; D006801:Humans; D007422:Intestines / Q000737:chemistry / Q000382:microbiology*; D048191:Lactiplantibacillus plantarum / Q000737:chemistry / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D019936:Probiotics / Q000737:chemistry / Q000378:metabolism; D000097573:Red Fluorescent Protein
chemical_list: D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein