OBJECTIVE: To clone and express the fused gene fragment coding rhoptry protein ROP2 and major surface protein P30 from Toxoplasma gondii as a preparation for the construction of the complex ROP2-P30 antigen by gene engineering. METHODS: The gene fragment encoding P30 was amplified by PCR from T. gondii RH strain and subcloned into the recombinant plasmid pUC119/ROP2 already constructed. The recombinant plasmid pUC119/ROP2-P30 was digested by Sac I/HindIII and inserted into the same site of expression vector pET28b. The recombinant plasmid of pET28b/ROP2-P30 was transformed to E. coli and expressed under the induction of IPTG. RESULTS: The gene fragment 700 bp encoding P30 was obtained from the total DNA of T. gondii by PCR. The recombinant plasmid pET28b/ROP2-P30 was successfully constructed, which was highly expressed in E. coli, a fusion protein with molecular weight of 69000. CONCLUSION: The fusion gene encoding the rhoptry protein ROP2 and the major surface protein P30 of T. gondii has been successfully cloned and expressed to be an expected recombinant fusion protein ROP2-P30 with molecular weight 69000.
OBJECTIVE: To clone and express the fused gene fragment coding rhoptry protein ROP2 and major surface protein P30 from Toxoplasma gondii as a preparation for the construction of the complex ROP2-P30 antigen by gene engineering. METHODS: The gene fragment encoding P30 was amplified by PCR from T. gondii RH strain and subcloned into the recombinant plasmid pUC119/ROP2 already constructed. The recombinant plasmid pUC119/ROP2-P30 was digested by Sac I/HindIII and inserted into the same site of expression vector pET28b. The recombinant plasmid of pET28b/ROP2-P30 was transformed to E. coli and expressed under the induction of IPTG. RESULTS: The gene fragment 700 bp encoding P30 was obtained from the total DNA of T. gondii by PCR. The recombinant plasmid pET28b/ROP2-P30 was successfully constructed, which was highly expressed in E. coli, a fusion protein with molecular weight of 69000. CONCLUSION: The fusion gene encoding the rhoptry protein ROP2 and the major surface protein P30 of T. gondii has been successfully cloned and expressed to be an expected recombinant fusion protein ROP2-P30 with molecular weight 69000.