Abstract: T. gondii producing proteins are strong antigens that can start strong immune reactions and one of these kinds of antigens is rhoptry protein 1 (ROP1) that is discharged from rhoptry cell-organ. These entire attribute for ROP1 makes it a competitor for protein vaccine and recombining vaccine against toxoplasmosis. A main objective of the current study was the Cloning and expression ROP1 of Toxoplasma gondii in a cloning vector for later studies. In the present work, genomic DNA of Toxoplasma gondii was removed and used for amplifying of ROP1 gene as a template. Then PCR product was cloned into the EcoR1 and BamH1 sites of cloning vector, pUET1 and transformed into Escherichia coli BL21 plysS strain and sub cloned from pTROP1 into the HindIII and EcoRI sites of the pcDNA3 to produce recombining eukaryotic declaration vector pcROP1. The cloned ROP1 was verified by PCR, limitation enzymes (HindIII and BglI) digestion and nucleotide sequencing. A fragment about 757 bp was separated fore more nucleotide sequence analysis of the ROP1 cloned in pUET1 vector revealed high homology (96%) with RH strain Gene Bank Accession No. M71274. This expressed ROP1 to make recombinant vaccine against toxoplasmosis will be useful in the future study.