title: Expression of Helicobacter pylori TonB protein in transgenic Arabidopsis thaliana: toward production of vaccine antigens in plants.
abstract: 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.
  
  
  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 administration as a route for immunization against Helicobacter infections.
authors.affiliation: Örebro Life Science Center, Örebro University, SE-70182 Örebro, Sweden.
authors.forename: Irina
authors.initials: I
authors.lastname: Kalbina
authors.forename: Lars
authors.initials: L
authors.lastname: Engstrand
authors.forename: Sören
authors.initials: S
authors.lastname: Andersson
authors.forename: Ake
authors.initials: A
authors.lastname: Strid
mesh_terms: D000942:Antigens, Bacterial / Q000096:biosynthesis* / Q000235:genetics; D017360:Arabidopsis / Q000235:genetics*; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D001428:Bacterial Vaccines / Q000096:biosynthesis* / Q000235:genetics; D015139:Blotting, Southern; D015153:Blotting, Western; D015870:Gene Expression; D018014:Gene Transfer Techniques; D016480:Helicobacter pylori / Q000235:genetics*; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012231:Rhizobium / Q000235:genetics
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D008565:Membrane Proteins; D011994:Recombinant Proteins; C057616:tonB protein, Bacteria