[Construction and verification of the dual-promoter plasmid pCN-SSISG as bivalent anti-caries DNA vaccine].
OBJECTIVE
The aim of the study is to construct an anti-caries DNA vaccine with high immunogenicity due to its unique properties of being able to express target antigens both in eukaryotic and prokaryotic host cells and harboring two main virulence genes from caries pathogen Streptoccocus mutans (S. mutans).


METHODS
The gene encoding glucan binding region (GBR) of glucosyltransferase-I (GTF-I) was amplified from the plasmid harboring the gtfB gene from S. mutans by PCR. Then the GBR gene that was upstream linked with tissue-type plasminogen activator signal peptide (tPA-SP) was directly cloned into the plasmid pCN-SSIE containing sSBR gene encoding tPA-SP and saliva binding region (SBR) of antigen protein I/II (AgI/II) from S. mutans. And finally the recombinant plasmid pCN-SSISG was analysed by DNA sequencing and endonuclearase digestion mapping.


RESULTS
DNA sequencing and endonuclearase digestion mapping showed that the open reading frame (ORF) and the tPA-SP sequence of the recombinant plasmid pCN-SSISG were identical with the expectant ones.


CONCLUSIONS
We have successfully constructed the dual-promoter bivalent recombinant plasmid pCN-SSISG.
Wang
Xiao-hua
XH
School of Dentistry, Shandong University, Jinan 250012, Shandong Province, China. wangxiaohua@mail.sdu.edu.cn
Jiang
Guang-shui
GS
Wu
Qin-zhen
QZ
Jiang
Ping-ping
PP
Liu
Hao
H
Jiang
Hao
H
D003731:Dental Caries / Q000517:prevention & control*; D005964:Glucosyltransferases / Q000235:genetics*; D010957:Plasmids; D012463:Saliva; D013295:Streptococcus mutans / Q000235:genetics; D019444:Vaccines, DNA
D019444:Vaccines, DNA; D005964:Glucosyltransferases; C489364:glucosyltransferase I