Cloning and Expression of Toxoplasma Gondii Major Surface Antigen 1 Truncated Fragment

To clone and express Toxoplasma gondii major surface antigen 1 truncated fragment and analyze its immunological activity.The gene encoding Toxoplasma gondii major surface antigen 1 truncated fragment was amplified by PCR from Toxoplasma gondii genomic DNA,and was cloned into pMD18 Tvector.Positive clones were identified by PCR and sequencing with ABIPRISMTM 377XL DNA sequencer;The insert digested with BamHⅠ and SalⅠwas subcloned into pGEX-4T-1 which was also digested with BamHⅠ and SalⅠ;There combinant plasmids were transformed into E.coli BL21 and were identified by PCR and double enzyme digestion;The recombinant clone was induced with IPTG to express target protein and characterized by SDS-PAGE and immunoblot.The results showed,the gene fragment encoding Toxoplasma gondii major surface antigen 1 truncated fragment was amplified by PCR from from Toxoplasma gondii genomic DNA.The inser of positive clone was subcloned into pGEX-4T-1 correctly;Recombinant protein was expressed in the positive recombinant clone when induced with IPTG and reacted with the rabbit sera infected with Toxoplasma gondii.The findings indicated that the recombinant plasmid expressing Toxoplasma gondii major surface antigen 1 truncated fragment was successfully constructed and the recombinant protein showed good immunologica lactivity.