1887

Abstract

Unlike , is unable to import a range of sugars, including arabinose, which makes common expression vectors, such as pBAD33, non-functional in these bacteria.

The aim of this study was to investigate whether the transporters AraE and modified LacY (LacYA177C) would enable to uptake arabinose.

The respective genes of were constitutively expressed in strain 11168H after integration into the chromosome via homologous recombination. Vectors carrying these genes also contained a reporter gene, , under the control of the arabinose-inducible promoter, pBAD. These constructs were verified in by demonstrating the induction of in the presence of arabinose. Integration of the genes into one of the rRNA gene clusters was verified by PCR and genome sequencing. The latter also confirmed that the inserted gene clusters contained no mutations. Expression of the gene in the presence of arabinose inducer was monitored using fluorescence microscopy of colonies and fluorimetry using both whole cells and lysates.

The results demonstrated the inability of to use arabinose transporters, which are fully functional in , suggesting a remarkable difference in the physiology of these bacteria.

  • This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Loading

Article metrics loading...

/content/journal/acmi/10.1099/acmi.0.000042
2019-07-01
2024-10-16
Loading full text...

Full text loading...

/deliver/fulltext/acmi/1/5/acmi000042.html?itemId=/content/journal/acmi/10.1099/acmi.0.000042&mimeType=html&fmt=ahah

References

  1. Bolton DJ. Campylobacter virulence and survival factors. Food Microbiol 2015; 48:99–108 [View Article]
    [Google Scholar]
  2. Byrne CM, Clyne M, Bourke B. Campylobacter jejuni adhere to and invade chicken intestinal epithelial cells in vitro. Microbiology 2007; 153:561–569 [View Article]
    [Google Scholar]
  3. Kaakoush NO, Castaño-Rodríguez N, Mitchell HM, Man SM. Global epidemiology of Campylobacter infection. Clin Microbiol Rev 2015; 28:687–720 [View Article]
    [Google Scholar]
  4. Altekruse SF, Stern NJ, Fields PI, Swerdlow DL. Campylobacter jejuni—an emerging foodborne pathogen. Emerg Infect Dis 1999; 5:28–35 [View Article]
    [Google Scholar]
  5. Gallardo F, Gascón J, Ruiz J, Corachan M, Jimenez de Anta M et al. Campylobacter jejuni as a cause of traveler's diarrhea: clinical features and antimicrobial susceptibility. J Travel Med 1998; 5:23–26 [View Article]
    [Google Scholar]
  6. Epps SV, Harvey RB, Hume ME, Phillips TD, Anderson RC et al. Foodborne Campylobacter: infections, metabolism, pathogenesis and reservoirs. Int J Environ Res Public Health 2013; 10:6292–6304 [View Article]
    [Google Scholar]
  7. Silva J, Leite D, Fernandes M, Mena C, Gibbs PA et al. Campylobacter spp. as a foodborne pathogen: a review. Front Microbiol 2011; 2:200 [View Article]
    [Google Scholar]
  8. Harvey P, Leach S. Analysis of coccal cell formation by Campylobacter jejuni using continuous culture techniques, and the importance of oxidative stress. J Appl Microbiol 1998; 85:398–404 [View Article]
    [Google Scholar]
  9. Jones DM, Sutcliffe EM, Curry A. Recovery of viable but non-culturable Campylobacter jejuni. J Gen Microbiol 1991; 137:2477–2482 [View Article]
    [Google Scholar]
  10. Signoretto C, Stefano FD, Canepari P. Modified peptidoglycan chemical composition in shape-altered Escherichia coli. Microbiology 1996; 142:1919–1926 [View Article]
    [Google Scholar]
  11. Amano K, Shibata Y. Structural Studies of Peptidoglycans in Campylobacter Species. Microbiol Immunol 1992; 36:961–967 [View Article]
    [Google Scholar]
  12. Costa K, Bacher G, Allmaier G, Dominguez-Bello M, Engstrand L et al. The morphological transition of Helicobacter pylori cells from spiral to coccoid is preceded by a substantial modification of the cell wall. J. Bacteriol 1999; 181:3710–3715
    [Google Scholar]
  13. Chaput C, Ecobichon C, Cayet N, Girardin SE, Werts C et al. Role of AmiA in the morphological transition of Helicobacter pylori and in Immune Escape. PLoS Pathog 2006; 2:e97–852 [View Article]
    [Google Scholar]
  14. Ikeda N, Karlyshev AV. Putative mechanisms and biological role of coccoid form formation in Campylobacter jejuni. Eur J Microbiol Immunol 2012; 2:41–49 [View Article]
    [Google Scholar]
  15. Newman JR, Fuqua C. Broad-host-range expression vectors that carry the L-arabinose-inducible Escherichia coli araBAD promoter and the araC regulator. Gene 1999; 227:197–203 [View Article]
    [Google Scholar]
  16. Fritz G, Megerle JA, Westermayer SA, Brick D, Heermann R et al. Single cell kinetics of phenotypic switching in the arabinose utilization system of E. coli. PLoS One 2014; 9:e89532 [View Article]
    [Google Scholar]
  17. Siegele DA, Hu JC. Gene expression from plasmids containing the araBAD promoter at subsaturating inducer concentrations represents mixed populations. Proc Natl Acad Sci USA 1997; 94:8168–8172 [View Article]
    [Google Scholar]
  18. Khlebnikov A, Risa O, Skaug T, Carrier TA, Keasling JD. Regulatable arabinose-inducible gene expression system with consistent control in all cells of a culture. J Bacteriol 2000; 182:7029–7034 [View Article]
    [Google Scholar]
  19. Khlebnikov A, Datsenko KA, Skaug T, Wanner BL, Keasling JD. Homogeneous expression of the P(BAD) promoter in Escherichia coli by constitutive expression of the low-affinity high-capacity AraE transporter. Microbiology 2001; 147:3241–3247 [View Article]
    [Google Scholar]
  20. Morgan-Kiss RM, Wadler C, Cronan JE. Long-term and homogeneous regulation of the Escherichia coli araBAD promoter by use of a lactose transporter of relaxed specificity. Proc Natl Acad Sci USA 2002; 99:7373–7377 [View Article]
    [Google Scholar]
  21. Karlyshev AV, Wren BW. Development and application of an insertional system for gene delivery and expression in Campylobacter jejuni. Appl Environ Microbiol 2005; 71:4004–4013 [View Article]
    [Google Scholar]
  22. Guzman LM, Belin D, Carson MJ, Beckwith J. Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J Bacteriol 1995; 177:4121–4130 [View Article]
    [Google Scholar]
  23. Zhang Y, Shang X, Lai S, Zhang G, Liang Y et al. Development and application of an arabinose-inducible expression system by facilitating inducer uptake in Corynebacterium glutamicum. Appl Environ Microbiol 2012; 78:5831–5838 [View Article]
    [Google Scholar]
  24. Parkhill J, Wren BW, Mungall K, Ketley JM, Churcher C et al. The genome sequence of the food-borne pathogen Campylobacter jejuni reveals hypervariable sequences. Nature 2000; 403:665–668 [View Article]
    [Google Scholar]
  25. Karlyshev AV, Linton D, Gregson NA, Wren BW. A novel paralogous gene family involved in phase-variable flagella-mediated motility in Campylobacter jejuni. Microbiology 2002; 148:473–480 [View Article]
    [Google Scholar]
  26. van Vliet A, Wooldridge K, Ketley J. Iron-responsive gene regulation in a Campylobacter jejuni fur mutant. J. Bacteriol 1998; 180:5291–5298
    [Google Scholar]
  27. Holt JP, Grant AJ, Coward C, Maskell DJ, Quinlan JJ. Identification of Cj1051c as a Major Determinant for the Restriction Barrier of Campylobacter jejuni Strain NCTC11168. Appl Environ Microbiol 2012; 78:7841–7848 [View Article]
    [Google Scholar]
/content/journal/acmi/10.1099/acmi.0.000042
Loading
/content/journal/acmi/10.1099/acmi.0.000042
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error