Transgenic carrot tap roots expressing an immunogenic F1-V fusion protein from Yersinia pestis are immunogenic in mice

J Plant Physiol. 2011 Jan 15;168(2):174-80. doi: 10.1016/j.jplph.2010.06.012. Epub 2010 Jul 23.

Abstract

Expression of the protective F1 and V antigens of Yersinia pestis, as a fusion protein, in carrot was pursued in an effort to develop an alternative vaccine production system against the serious plague disease. Transgenic carrot plants carrying the F1-V encoding gene were developed via Agrobacterium-mediated transformation. Presence, integration, and expression of the F1-V encoding gene were confirmed by polymerase chain reaction (PCR), DNA gel blot analysis, and reverse-transcriptase (RT)-PCR analyses, respectively. An ELISA assay confirmed the antigenicity of the plant-derived F1-V fusion protein. Immunogenicity was evaluated subcutaneously in mice using a soluble protein extract of freeze-dried transgenic carrot. Significant antibody levels were detected following immunization. These results demonstrated that the F1-V protein could be expressed in carrot tap roots, and that the carrot F1-V recombinant protein retained its antigenicity and immunogenicity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Bacterial / genetics
  • Antigens, Bacterial / metabolism*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Daucus carota / genetics
  • Daucus carota / metabolism*
  • Enzyme-Linked Immunosorbent Assay
  • Mice
  • Mice, Inbred BALB C
  • Plague / immunology
  • Plague Vaccine / genetics
  • Plague Vaccine / immunology
  • Plague Vaccine / metabolism
  • Plant Roots / genetics
  • Plant Roots / metabolism*
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / metabolism*
  • Pore Forming Cytotoxic Proteins / genetics
  • Pore Forming Cytotoxic Proteins / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Yersinia pestis / genetics
  • Yersinia pestis / metabolism*

Substances

  • Antigens, Bacterial
  • Bacterial Proteins
  • LcrV protein, Yersinia
  • Plague Vaccine
  • Pore Forming Cytotoxic Proteins
  • Recombinant Fusion Proteins
  • caf1 protein, Yersinia pestis