Toxoplasma gondii SAG1,SAG3 Cloning and composite SAG1/SAG3 gene expression of recombinant plasmid

Objective To construct eukaryotic expression recombinant of surface antigen SAG1 and SAG3 of Toxoplasma,preparate for further toxoplasmosis vaccine study.Methods extract the Toxoplasma's genome DNA;amplify the surface antigen SAG1 and SAG3 genes by PCR technology,link two genes to pGEM-T cloning vector respectively;after enzyme digestion and purification take two genes subclone into pcDNA3.1(+) eukaryotic expression vector,then by PCR、enzyme digestion and sequencing methods identify the recombinant plasmid.Results amplified SAG1 and SAG3 genes from toxoplasmosis genome;successfully constructed pGEM-SAG1 and pGEM-SAG3 cloning plasmids;constructed pcDNA3.1-SAG1-SAG3 complex gene expression vector,DNA sequencing showed that connect the right direction.Conclusion constructed pcDNA3.1-SAG1-SAG3 compound gene expression vector,to provide the candidate antigen and lay the experiment foundation for the next development Toxoplasmosis compound multiple vaccine.