Construction of a targeted DNA vaccine against periodontitis and its expression in vitro

Objective:To construct a CTLA4 targeted DNA vaccine against FimA of Porphyromonas gingivalis and detect its expression in vitro.Method:The CTLA4-Ig gene and fimA gene were amplified from plasmid pGJGLU/GFP and pcDNA3/fimA respectively.The targeted plasmid pCTLA4-FimA was constructed by cloning these two fragments into pCI vector.COS7 cells were transfected with pCTLA4-FimA and the expression of proteins were detected by Western blotting and indirect immunofluorescence.Result:pCTLA4-FimA contains both CTLA4-Ig gene and fimA gene.The expressed protein could react to specific anti-FimA antibody.Green fluorescence was detected in the cytoplasm of pCTLA4-FimA transfected CHO cells incubated with anti-FimA antibody.Conclusion:The targeted DNA vaccine pCTLA4-FimA is constructed successfully and can be expressed correctly in COS7 cells in vitro.