Construction of Targeted DNA Vaccine against Glucosyltransferase-I of Streptococcus mutans

Objectives: To construct and express in vitro the targeted anti-caries DNA vaccine pGJGLU encoding the glu fragment of gtfB and extracellular region of the human CTLA4 for the targeting antigen to APC. Methods: The A-P fragment of pGJA-P, a targeted anti-caries DNA vaccine encoding extracellular region of the human CTLA4, Fc region of human ¦Ã1, the glu fragment of gtfB and A-P fragment of pac gene of Streptococcus mutans, was deleted. A stop codon was added to the downstream of glu fragment to construct the targeted anti-caries DNA vaccine pGJGLU. CHO cells were transfected with pGJGLU by liposome. The expression of recombinant protein in cultured supernatant or in cells was detected using Western blotting or cellular immunofluorescence respectively. Results: pGJGLU carried the fusion gene of CTLA4-Ig fusion gene and glu fragment of gtfB gene. The expressed recombinant protein could response to specific anti-GTF antibody. The recombinant protein can be detected both in cultured supernatant of CHO cells and inside the CHO cells transfected with pGJGLU. The recombinant protein existed as a dimer under non-reducing conditions. Conclusions: The targeted anti-caries DNA vaccine pGJGLU was constructed successfully and expressed the recombinant protein in eukaryotic cells correctly. The study was supported by National Natural Scientific Foundation of China. 30271416