This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Mixter, P. F.
Right arrow Articles by Konkel, M. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mixter, P. F.
Right arrow Articles by Konkel, M. E.
Agricola
Right arrow Articles by Mixter, P. F.
Right arrow Articles by Konkel, M. E.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, May 2003, p. 2864-2874, Vol. 69, No. 5
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.5.2864-2874.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

In Vivo Tracking of Campylobacter jejuni by Using a Novel Recombinant Expressing Green Fluorescent Protein

Philip F. Mixter, John D. Klena, Gary A. Flom, Amy M. Siegesmund, and Michael E. Konkel*

School of Molecular Biosciences, Washington State University, Pullman, Washington 99164-4234

Received 4 October 2002/ Accepted 7 February 2003

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.

* Corresponding author. Mailing address: School of Molecular Biosciences, Washington State University, Pullman, WA 99164-4234. Phone: (509) 335-5039. Fax: (509) 335-1907. E-mail: konkel{at}mail.wsu.edu.

Applied and Environmental Microbiology, May 2003, p. 2864-2874, Vol. 69, No. 5
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.5.2864-2874.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Dothager, R. S., Piwnica-Worms, D. (2009). Molecular Imaging of Pulmonary Disease In Vivo. Proc Am Thorac Soc 6: 403-410 [Abstract] [Full Text]  
  • Al-Banna, N. A., Junaid, T. A., Mathew, T. C., Raghupathy, R., Albert, M. J. (2008). Histopathological and ultrastructural studies of a mouse lung model of Campylobacter jejuni infection. J Med Microbiol 57: 210-217 [Abstract] [Full Text]  
  • Malik-Kale, P., Raphael, B. H., Parker, C. T., Joens, L. A., Klena, J. D., Quinones, B., Keech, A. M., Konkel, M. E. (2007). Characterization of Genetically Matched Isolates of Campylobacter jejuni Reveals that Mutations in Genes Involved in Flagellar Biosynthesis Alter the Organism's Virulence Potential. Appl. Environ. Microbiol. 73: 3123-3136 [Abstract] [Full Text]  
  • Konkel, M. E., Christensen, J. E., Dhillon, A. S., Lane, A. B., Hare-Sanford, R., Schaberg, D. M., Larson, C. L. (2007). Campylobacter jejuni Strains Compete for Colonization in Broiler Chicks. Appl. Environ. Microbiol. 73: 2297-2305 [Abstract] [Full Text]  
  • Besser, T. E., Shaikh, N., Holt, N. J., Tarr, P. I., Konkel, M. E., Malik-Kale, P., Walsh, C. W., Whittam, T. S., Bono, J. L. (2007). Greater Diversity of Shiga Toxin-Encoding Bacteriophage Insertion Sites among Escherichia coli O157:H7 Isolates from Cattle than in Those from Humans. Appl. Environ. Microbiol. 73: 671-679 [Abstract] [Full Text]  
  • Karlyshev, A. V., Wren, B. W. (2005). Development and Application of an Insertional System for Gene Delivery and Expression in Campylobacter jejuni. Appl. Environ. Microbiol. 71: 4004-4013 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»