Journal article
In Vivo Tracking of Campylobacter jejuni by Using a Novel Recombinant Expressing Green Fluorescent Protein
Applied and environmental microbiology, Vol.69(5), pp.2864-2874
05/2003
Handle:
https://hdl.handle.net/2376/105385
PMCID: PMC154531
PMID: 12732559
Abstract
Campylobacter jejuni
is a leading cause of food-borne disease in developed countries. The goal of this study was to develop a plasmid-based reporter system with green fluorescent protein (GFP) to facilitate the study of
C. jejuni
in a variety of niches.
C. jejuni
transformants harboring the pMEK91 GFP gene (
gfp
)-containing vector were readily detectable by both fluorescence microscopy and flow cytometry. Given the ease of detecting these organisms, additional experiments were performed in which BALB/c mice were injected intraperitoneally with
C. jejuni
harboring the
gfp
-containing vector. Four hours after injection of the mice, flow cytometry analyses determined that
C. jejuni
synthesizing GFP were predominantly associated with granulocytes. More specifically, the proportion of CD11b
+
Gr-1
+
lavage neutrophils with green fluorescence ranged from 99.7 to 100%, while the proportion of CD11b
+
Gr-1
−
lavage macrophages ranged from 77.0 to 80.0%. In contrast, few CD11b
−
CD45R
+
B lymphocytes from the lavage of the
C. jejuni
-injected mice were associated with green-fluorescent
C. jejuni
(proportions ranged from 0.75 to 0.77%). Cell-free
C. jejuni
was recovered from tissue homogenates after intraperitoneal injection. Macrorestriction profiling with pulsed-field gel electrophoresis identified a genotypic variant of the
C. jejuni
F38011 wild-type isolate. In vivo this variant displayed a phenotype identical to that of the wild-type isolate. In summary, we demonstrate that
C. jejuni
associates with marker-defined cellular subsets in vivo with a novel
gfp
reporter system and that
C. jejuni
genotypic variants can be isolated from both in vitro and in vivo systems.
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Details
- Title
- In Vivo Tracking of Campylobacter jejuni by Using a Novel Recombinant Expressing Green Fluorescent Protein
- Creators
- Philip F Mixter - School of Molecular Biosciences, Washington State University, Pullman, Washington 99164-4234John D Klena - School of Molecular Biosciences, Washington State University, Pullman, Washington 99164-4234Gary A Flom - School of Molecular Biosciences, Washington State University, Pullman, Washington 99164-4234Amy M Siegesmund - School of Molecular Biosciences, Washington State University, Pullman, Washington 99164-4234Michael E Konkel - School of Molecular Biosciences, Washington State University, Pullman, Washington 99164-4234
- Publication Details
- Applied and environmental microbiology, Vol.69(5), pp.2864-2874
- Academic Unit
- Molecular Biosciences, School of
- Publisher
- American Society for Microbiology
- Identifiers
- 99900546805601842
- Language
- English
- Resource Type
- Journal article