Potato virus X coat protein fusion to human papillomavirus 16 E7 oncoprotein enhance antigen stability and accumulation in tobacco chloroplast

Mol Biotechnol. 2009 Nov;43(3):243-9. doi: 10.1007/s12033-009-9195-3. Epub 2009 Jul 5.

Abstract

Cervical cancer linked to infection with human papillomavirus (HPV) is the third cause of cancer-related death in women. As the virus cannot be propagated in culture, vaccines have been based on recombinant antigens with inherited high-cost production. In a search of alternative cheap production system, E7 HPV type 16 protein, an attractive candidate for anticancer vaccine development, was engineered to be expressed in tobacco chloroplast. In addition, E7 coding sequence was fused to potato virus X coat protein (CP) to compare expression level. Results show that E7CP transcript accumulation reached lower levels than non-fused E7. However, antigen expression levels were higher for fusion protein indicating that CP stabilizes E7 peptide in the chloroplast stroma. These results support viability of transplastomic plants for antigen production and the relevance of improving recombinant peptide stability for certain transgenes to enhance protein accumulation in this organelle.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Northern
  • Blotting, Southern
  • Capsid Proteins / genetics*
  • Capsid Proteins / metabolism
  • Chloroplasts / genetics*
  • Chloroplasts / metabolism
  • Cloning, Molecular
  • Nicotiana / genetics*
  • Nicotiana / metabolism
  • Oncogene Proteins, Viral / genetics*
  • Oncogene Proteins, Viral / metabolism
  • Papillomavirus E7 Proteins
  • Plants, Genetically Modified / genetics*
  • Plants, Genetically Modified / metabolism
  • Polymerase Chain Reaction
  • Protein Stability
  • Recombinant Fusion Proteins / genetics*
  • Recombinant Fusion Proteins / metabolism

Substances

  • Capsid Proteins
  • Oncogene Proteins, Viral
  • Papillomavirus E7 Proteins
  • Recombinant Fusion Proteins
  • coat protein, Potato virus X
  • oncogene protein E7, Human papillomavirus type 16