Construction of GM-CSF-Survivin Fusion Gene and Its Expression in Dendritic Cells

Objective: To construct GM-CSF-Survivin fusion gene with Survivin as tumor antigen and GM-CSF as adjuvant gene and transfect it into dendritic cells.To study new method of anti-tumor effect of DC vaccine.Methods: GM-CSF-Survivin DNA fragments were obtained by PCR reaction and connected with pcDNA3.1(-) plasmid vector.Transform the conjugate into Ecoli DH5α,select clone and extract plasmid.The plasmid was transfected into DC by jetPEITM-Macrophage systerm.Use PCR and Western blot to identify the recombinating plasmid.Results: GM-CSF-Survivin mRNA and protein were overexpressed in DC.Conclusion: We have constructed pcDNA3.1(-)-GM-CSF-CSF-Survivin successfully and transfected it into DC.