AI Article Synopsis

  • The study explores using bacterial endospores as a system to display antigens, focusing on the UreA subunit of urease from Helicobacter acinonychis, a pathogen linked to virulence in bacteria.
  • The research involves expressing UreA on the Bacillus subtilis spore coat using different proteins (CotB, CotC, CotG) as carriers and comparing their efficiency in surface expression and display.
  • Results indicate that UreA fused to CotB is effectively displayed on the spore surface, while CotC allows the highest expression rate but does not present UreA on the surface, making CotB the best choice for displaying foreign proteins.

Article Abstract

Background: The bacterial endospore (spore) has recently been proposed as a new surface display system. Antigens and enzymes have been successfully exposed on the surface layers of the Bacillus subtilis spore, but only in a few cases the efficiency of expression and the effective surface display and have been determined. We used this heterologous expression system to produce the A subunit of the urease of the animal pathogen Helicobater acinonychis. Ureases are multi-subunit enzymes with a central role in the virulence of various bacterial pathogens and necessary for colonization of the gastric mucosa by the human pathogen H. pylori. The urease subunit UreA has been recognized as a major antigen, able to induce high levels of protection against challenge infections.

Results: We expressed UreA from H. acinonychis on the B. subtilis spore coat by using three different spore coat proteins as carriers and compared the efficiency of surface expression and surface display obtained with the three carriers. A combination of western-, dot-blot and immunofluorescence microscopy allowed us to conclude that, when fused to CotB, UreA is displayed on the spore surface (ca. 1 x 10(3) recombinant molecules per spore), whereas when fused to CotC, although most efficiently expressed (7-15 x 10(3) recombinant molecules per spore) and located in the coat layer, it is not displayed on the surface. Experiments with CotG gave results similar to those with CotC, but the CotG-UreA recombinant protein appeared to be partially processed.

Conclusion: UreA was efficiently expressed on the spore coat of B. subtilis when fused to CotB, CotC or CotG. Of these three coat proteins CotC allows the highest efficiency of expression, whereas CotB is the most appropriate for the display of heterologous proteins on the spore surface.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2841587PMC
http://dx.doi.org/10.1186/1475-2859-9-2DOI Listing

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