ApiLoc - A database of published protein sub-cellular localisation in Apicomplexa

version 3 (curated until May 28, 2011)

Toxoplasma gondii: cloning, sequencing, expression, and antigenic characterization of ROP2, GRA5 and GRA7.

Igarashi, M., Kano, F., Tamekuni, K., Kawasaki, P. M., Navarro, I. T., Vidotto, O., Vidotto, M. C., Machado, R. Z., Garcia, J. L. (2008, Genet Mol Res)

Toxoplasma gondii is an intracellular obligate protozoan, which infects humans and warm-blooded animals. The aim of the present study was to clone the rop2, gra5 and gra7 genes from T. gondii RH strain and to produce recombinant proteins. The rop2, gra5 and gra7 gene fragments produced by polymerase chain reaction were cloned into the pET102/D-TOPO vector which contains thioredoxin and polyhistidine tags at the C- and N-ends, respectively, and is expressed in Escherichia coli BL21(DE-3). The expression fusion proteins were found almost entirely in the insoluble form in the cell lysate. These recombinant proteins were purified with an Ni-NTA column. Concentrations of the recombinant antigens produced in the E. coli BL21-star ranged from 300 to 500 microg/ml growth media, which was used to immunize rabbits. We observed an identity ranging from 96 to 97% when nucleotide sequences were compared to GenBank database sequences. Immunocharacterization of proteins was made by indirect immunofluorescence assay. These proteins will be used for serodiagnosis and vaccination.

PubMed: 18551396, full text

Localisation information

TGME49_015780 (ROP2/3/4, ROP2, Tg3, Tg4, Tg34) Rhoptry kinase family protein ROP2A (incomplete catalytic triad)

Experimental localisation: rhoptry during tachyzoite
  • Species: Toxoplasma gondii
  • Quote inferring localisation: "The present study cloned genes from two secretory organelles from T. gondii, dense granules and rhoptry. Analysis of sequence variation in the T. gondii rop2-, gra5- and gra7- coding regions from Londrina-RH strain showed some heterogeneity in nucleotides and predicted amino acid sequences (data not shown), when compared to the RH strains from the GenBank. However, this heterogeneity neither changed reactivity in the indirect fluorescence antibody test nor immunoblotting to T. gondii tachyzoites."
  • Microscopy type: Light
  • Microscopy method: antibody directly to protein
  • Strain: RH
  • Gene model mapping comments: Blast from Z36906, annotation matches
  • Localisation record: rhoptry during tachyzoite

TGME49_086450 (GRA5) dense granule protein 5 precursor

Experimental localisation: dense granule during tachyzoite
  • Species: Toxoplasma gondii
  • Quote inferring localisation: "The present study cloned genes from two secretory organelles from T. gondii, dense granules and rhoptry. Analysis of sequence variation in the T. gondii rop2-, gra5- and gra7- coding regions from Londrina-RH strain showed some heterogeneity in nucleotides and predicted amino acid sequences (data not shown), when compared to the RH strains from the GenBank. However, this heterogeneity neither changed reactivity in the indirect fluorescence antibody test nor immunoblotting to T. gondii tachyzoites."
  • Microscopy type: Light
  • Microscopy method: antibody directly to protein
  • Strain: RH
  • Gene model mapping comments: Blast from L06091, annotation matches
  • Localisation record: dense granule during tachyzoite

TGME49_003310 (GRA7) dense granule protein 7

Experimental localisation: dense granule during tachyzoite
  • Species: Toxoplasma gondii
  • Quote inferring localisation: "The present study cloned genes from two secretory organelles from T. gondii, dense granules and rhoptry. Analysis of sequence variation in the T. gondii rop2-, gra5- and gra7- coding regions from Londrina-RH strain showed some heterogeneity in nucleotides and predicted amino acid sequences (data not shown), when compared to the RH strains from the GenBank. However, this heterogeneity neither changed reactivity in the indirect fluorescence antibody test nor immunoblotting to T. gondii tachyzoites."
  • Microscopy type: Light
  • Microscopy method: antibody directly to protein
  • Strain: RH
  • Gene model mapping comments: Blast from Y13863, annotation matches
  • Localisation record: dense granule during tachyzoite