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Expression of Helicobacter pylori TonB Protein in Transgenic Arabidopsis thaliana: toward production of vaccine antigens in plants
Örebro University, School of Science and Technology.
Dept Bacteriol, Swedish Inst Infect Dis Control SMI, Solna, Sweden.
Örebro University Hospital, Örebro, Sweden; Dept Bacteriol, Swedish Inst Infect Dis Control SMI, Solna, Sweden.ORCID iD: 0000-0002-4522-3078
Örebro University, School of Science and Technology.ORCID iD: 0000-0003-3315-8835
2010 (English)In: Helicobacter, ISSN 1083-4389, E-ISSN 1523-5378, Vol. 15, no 5, p. 430-437Article in journal (Refereed) Published
Abstract [en]

Background: The aim of this study was to produce a recombinant version of the highly antigenic Helicobacter pylori TonB (iron-dependent siderophore transporter protein HP1341) in transgenic plants as a candidate oral vaccine antigen. Materials and Methods: Using Agrobacterium-mediated gene transfer, we introduced three different constructs of the tonB gene into the genome of the model plant Arabidopsis thaliana. We investigated transgene insertion by PCR, produced TonB antibodies for analysis of the production of the recombinant protein in plants, verified the identity of the protein produced by mass spectrometry analysis, and analyzed the number of genetic inserts in the plants by Southern blotting. Results: Three different constructs of the expression cassette (full-length tonB, tonB truncated in the 5' end removing the codons for a transmembrane helix, and the latter construct with codons for the endoplasmic reticulum SEKDEL retention signal added to the 3' end) were used to find the most effective way to express the TonB antigen. Production of TonB protein was detected in plants transformed with each of the constructs, confirmed by both Western blotting and mass spectrometry analysis. No considerable differences in protein expression from the three different constructs were observed. The protein concentration in the plants was at least 0.05% of the total soluble proteins. Conclusions: The Helicobacter pylori TonB protein can be produced in Arabidopsis thaliana plants in a form that is recognizable by rabbit anti-TonB antiserum. These TonB-expressing plants are highly suitable for animal studies of oral adminstration as a route for immunization against Helicobacter infections.

Place, publisher, year, edition, pages
John Wiley & Sons, 2010. Vol. 15, no 5, p. 430-437
Keywords [en]
edible vaccines, Helicobacter pylori, HP1341, subunit vaccine, TonB, transgenic plants
National Category
Biochemistry and Molecular Biology
Research subject
Biochemistry
Identifiers
URN: urn:nbn:se:oru:diva-12889DOI: 10.1111/j.1523-5378.2010.00786.xISI: 000281636600004PubMedID: 21083749OAI: oai:DiVA.org:oru-12889DiVA, id: diva2:383450
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This is a pre-print article.

Available from: 2012-08-06 Created: 2011-01-03 Last updated: 2018-04-23Bibliographically approved

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Kalbina, IrinaAndersson, SörenStrid, Åke

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