Expression of complete rhoptry protein 2 (ROP2) gene of Toxoplasma gondii in eukaryotic cell

  • KH Khosroshahi
  • F Ghaffarifar
  • Z Sharifi
  • A Dalimi
Keywords: Cloning, sequencing, Toxoplasma gondii, ROP2

Abstract

Toxoplasma gondii is the intracellular protozoan parasite responsible for animal and human toxoplasmosis. In immunodeficient patients, chronic infection with T. gondii can reactivate and produce
encephalitis, which is often lethal. ROP2 (rhoptry protein of T. gondii) is one of the most important interferer in organelle and PVM blending. ROP2 protein is recognized by clone T-cell (Tcc32) in human
body and also has epitope for B-cell. All of these characteristics of ROP2 makes it a candidate for cocktail vaccine and recombinant vaccine against toxoplasmosis. We described the expression of the
gene which encodes the complete rhoptry protein 2 (ROP2) of T. gondii in CHO cells and confirmed it by SDS-PAGE and Western blot analysis. In the present work, genomic DNA of T. gondii was extracted
and used for amplifying of ROP2 gene as a template. Then PCR product was cloned into pTZ57R/T vector, and plasmid containing ROP2 gene (pT-ROP2) was extracted from transformed bacteria and
sequenced. We hope to use from this recombinant plasmid (pT-ROP2) to make DNA vaccine against toxoplasmosis.
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eISSN: 1684-5315