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Gene encoding GRA1 protein is potent DNA-vaccine candidate against toxoplasmosis. The aim of the research<br />was to clone the gene encoding GRA1 protein of tachyzoite Toxoplasma gondii local isolate by DNA recombinant<br />technology. Tachyzoite was grown in Balb/c mice in vivo. Messenger RNA was isolated from total RNA and it<br />was used to synthesis cDNA. Complementary DNA encoding GRA1 protein of tachyzoite Toxoplasma gondii local<br />isolate was amplified and cloned in a prokaryote cloning vector. The recombinant GRA1-encoding gene was then<br />digesting using EcoRI restriction endonuclease and sequencing. The result showed that the recombinant GRA1-<br />encoding gene consisted of DNA sequences encoding all signal peptide and mature peptide of GRA1 protein.<br />Alignment of recombinant GRA1 sequence to gene encoding GRA1 protein of Toxoplasma gondii RH isolate showed<br />100% homologous.<br />Keywords: GRA1 protein, Toxoplasma gondii, tachyzoite, cloning, cDNA