Toxoplasma Full-Length cDNA Project (T.gondii)
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Last Updated (October 15, 2004)

About this Database
Toxoplasma Full-Length cDNA project



2.Construction of Toxoplasma Full-Length cDNA Library


I) Cultivation of Toxoplasma parasites

The RH strain of Toxoplasma gondii tachyzoites (1x107 parasites) were infected to Vero (T25) cells and incubated in EMEM medium supplemented with 8% fetal bovine serum for 72 hours in vitro.
Infected cells were collected with a scraper and passed through 27G needle for three times and purified by filtration through Millex SV filters (pore size 5 um, Millipore).
Cells were collected by centrifugation at 2,000 rpm for 10 min and washed in EMEM medium without FBS.
Parasites were suspended at 1x107/ml in EMEM medium without FBS. Five-week old ICR female mice were infected with 1 ml parasites i.p..
Three days after infection, mice were sacrificed by cervical dislocation. Parasites were collected by washing the peritoneal cavity with 5 ml EMEM, passed through 5 um filter, washed in PBS and suspended at 2-3x109 parasite/ml.
20 ml TRI REGENT (Sigma) was added to 1 ml parasite solution, mixed by pipetting, frozen in liquid nitrogen and kept at -80C until used.

II) Production of a full-length cDNA

A full-length enriched library was produced by oligo-capping method as described in Full-malaria.


  1. Toxoplasma
  2. Construction of Toxoplasma Full-Length cDNA Library
  3. Current Status

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