Construction and eukaryotic expression of genetic vaccine encoding G250 complex antigen for renal cell carcinoma

Objective: To construct a eukaryotic expression plasmid of tG250 fusion genes encoding the most Cytotoxic T Lymphocyte Epitopes of human G250 and part of G250 genes of mice and monkeys,and detect the expression in the eukaryotic cell COS7. Methods: tG250 fusion genes were constructed by gene synthesis and PCR.tG250 genes were inserted into a eukaryotic expression vector pCI-Fc-GPI that included the genes of the targeting signal peptide of human Igk,human IgG-Fc and GPI.The recombinant plasmid PCI-Fc-tG250 GPI was transfected to the COS7 cells,and the expression was detected by FACS（Fluorescence Activated Cell Sorting）and IMF（immunofluorescence）. Results: The sequence of tG250 fusion genes was consistent with the design.PCR and enzyme digestion analysis showed that the recombinant plasmid PCI-Fc-tG250-GPI was successfully constructed.The expression of this plasmid was demonstrated by FACS and IMF. Conclusion: The recombinant plasmid pCI-Fc-tG250-GPI has been successfully constructed and expressed in the COS7 cells.These results have provided necessary bases for the study of the anti-tumor effects of this cancer vaccine targeting G250 in the future.