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Potato virus X displaying the E7 peptide derived from human papillomavirus type 16: a novel position for epitope presentation

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Abstract

Potato virus X (PVX) is widely used as a peptide presentation system in plant biotechnology, mostly by fusion of desired peptides to the N-terminus of its capsid protein (CP). Considering that some epitopes can interfere with the stability and/or self-assembly of PVX CP when fused to its N-terminus, we evaluated four other possible sites for fusion using the E7 epitope derived from human papillomavirus type 16 with different tags. We prepared eight different PVX CP constructs modified with the E7 epitope fused with the 6xHis tag in both orientations (6xHis-E7, E7-6xHis), cloned them into the PVX-based vector pGR106 and expressed them transiently in Nicotiana benthamina plants. Only the fusion site located after amino acid 23 led to systemic infection of plants and the production of recombinant proteins, but no viral particles were detected. When we replaced the 6xHis with StrepII tag, the modified virus infected plants systemically, expressed proteins assembled into viral particles and the epitopes were located on the particle surface. The results of this study indicate that the new position within the PVX CP can be used for peptide presentation on the surface of PVX particles and is promising for PVX based production of therapeutic compounds in plants.

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Acknowledgments

This research was supported by the Grant No. P501/12/1761 of the Czech Science Foundation, CZ.2. 16/3.1.00/24014 from the European Regional Development Fund, Operational Programme Prague-Competitiveness and Grant No. 631412 of the Charles University Grant Agency. We acknowledge the skilled technical assistance of Mrs. R. Hadamkova and D. Cibochova.

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Correspondence to Noemi Cerovska.

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Vaculik, P., Plchova, H., Moravec, T. et al. Potato virus X displaying the E7 peptide derived from human papillomavirus type 16: a novel position for epitope presentation. Plant Cell Tiss Organ Cult 120, 671–680 (2015). https://doi.org/10.1007/s11240-014-0634-x

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Keywords

  • Potato virus X
  • Human papillomavirus
  • Nicotiana benthamiana
  • E7 oncoprotein
  • Transient expression