Efficient expression of a Toxoplasma gondii dense granule Gra4 antigen in tobacco leaves

Exp Parasitol. 2008 Sep;120(1):118-22. doi: 10.1016/j.exppara.2008.06.002. Epub 2008 Jun 10.

Abstract

A His-tagged truncated version of Toxoplasma gondii dense granule 4 protein (Gra4(163-345)) was transiently expressed in tobacco leaves. Two genetic constructions were used to accomplish this goal. In one of them, based in a Potato virus X (PVX) amplicon, the sequence encoding His-Gra4(163-345) was placed under control of an additional PVX coat protein subgenomic promoter. In the other, the same sequence was fused to an apoplastic transport signal and placed under the direction of the cauliflower mosaic virus 35S promoter. His-Gra4(163-345) accumulation in agroinfiltrated tobacco leaves was estimated by Western blot analysis using mouse anti-Gra4 antibody and a seropositive human serum. Here, we demonstrated the feasibility of producing a Gra4 antigen using transient expression methods in plants.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agrobacterium tumefaciens
  • Animals
  • Antigens, Protozoan / biosynthesis*
  • Antigens, Protozoan / genetics
  • Antigens, Protozoan / immunology
  • Blotting, Western
  • Caulimovirus
  • Gene Expression Regulation, Plant*
  • Genetic Vectors
  • Nicotiana / genetics
  • Nicotiana / metabolism*
  • Plant Leaves / genetics
  • Plant Leaves / metabolism
  • Protozoan Proteins / biosynthesis*
  • Protozoan Proteins / genetics
  • Protozoan Proteins / immunology
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Toxoplasma / genetics
  • Toxoplasma / immunology*

Substances

  • Antigens, Protozoan
  • GRA4 protein, Toxoplasma gondii
  • Protozoan Proteins
  • Recombinant Proteins