Construction And Expression Of Nucleic Acid Vaccine Of Human A-SYN Epitope Gene PVAX1-IL-4/SYN-B

Objective: To construct eukaryotic expression plasmid carryed human IL-4 gene and the B cell epitopes gene ofα-synuclein protein ,and investigate the expression of the plasmid in COS-7 cells. Methods :The gene of human IL-4 was obtained by RT-PCR and then was fused with the B cell epitopes gene ofα-synuclein protein. A short flexibility peptide was inserted(GSGGGSG)between the gene of human IL-4 and B cell epitomes. The fusional product was cloned into an eukaryotic expression plasmid pVAX1.Then the fusional plasmid was confirmed by electrophoresis and DNA sequencing. The fusional plasmid was transfected into COS-7 cells by liposome transfecting technique,and then identified by Western blotting.Results : The fusional plasmid,named pVAX1-IL-4 /SYN–B,was constructed and confirmed by DNA sequencing.Meanwhile,the protein extracting in COS-7 cells transfected with the plasmid was showed reactivity with antibody againstα-synuclein. Conclusion :The pVAX1-IL-4 /SYN–B was constructed successfully and highly expressed in COS-7 cells ,which lay a foundation for further studies.