[Construction of expression vector containing S. mutans GBR gene and confirmation of the immunogenicity of the recombinant GBR protein].
OBJECTIVE
The purpose of this study is to construct the expressing plasmid vector containing the GBR gene of Streptococcus mutans and study the immunogenicity of recombinant GBR protein.


METHODS
The GBR gene was cloned into the expression vector pTriEx-4 through gene cloning techniques by PCR,T-A clone and etc,the recombinant plasmid pTriEx-4-GBR was analyzed by DNA sequencing and endonuclearase digestion mapping. The recombinant GBR(rGBR) protein expression was induced with IPTG in E. coli JM109(DE3) which was transformed with plasmid pTriEx-4-GBR and then the rGBR protein was purified by affinity chromatography. Balb/c mice were randomly divided into two groups:mice of experimental group were inoculated subcutaneously with the rGBR protein and the rGBR protein was replaced for NS in the control group, then the anti-serum was evaluated by ELISA.The data were analysed by statistical software PEMS3.0.


RESULTS
The DNA sequence and the reading frame of GBR gene in the reconstructed vector pTriEx-4-GBR was in corresponding with the initial design and the rGBR protein was purified. The level of specific anti-rGBR serum IgG in the experimental group was significantly higher than that in the control group(P<0.005).


CONCLUSIONS
The results showed that the expressing plasmid carrying the GBR gene was constructed successfully and the purified recombinant GBR protein can elicit specific murine system immune response, which is necessary for further experiments including construction of bivalent anti-caries DNA vaccine and studies both in vitro and in vivo.
Wang
Xiao-hua
XH
School of Stomatology, Shandong University, Jinan 250012, Shandong Province, China.
Jiang
Guang-shui
GS
Wu
Qin-zhen
QZ
Jiang
Ping-ping
PP
Geng
Zhao
Z
Zhang
Bing
B
D000818:Animals; D003001:Cloning, Molecular; D003731:Dental Caries / Q000517:prevention & control; D004926:Escherichia coli; D005798:Genes, Bacterial; D005822:Genetic Vectors; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000235:genetics* / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics* / Q000276:immunology*; D013295:Streptococcus mutans / Q000235:genetics*
D011994:Recombinant Proteins