[Construction and expression in vitro of a targeted fusion anticaries DNA vaccine].
OBJECTIVE
To construct and detect the targeted anti-caries fusion DNA vaccine pGJA-P encoding the A-P fragment of pac, glu fragment of gtfB and extracellular region of the human CTLA4 and the Fc region of human Iggamma(1) gene for the targeting antigen to APC.


METHODS
The extracellular region of the human CTLA4 and the Fc region of human Iggamma(1) genes were amplified by RT-PCR from human lymphocytes, and the genes were cloned into pUC(m-T) vector respectively. After sequencing, Fc region of human Iggamma(1) gene was cloned to the downstream of CTLA4 gene fragment as the recombinant plasmid pGJ. The fusion gene was then inserted into the plasmid pGLUA-P to get the recombinant plasmid pGJA-P. The CHO cells were transfected with liposome and the expression of fusion protein in cultured supernatants were detected using Western blotting.


RESULTS
The plasmids pGJ and pGJA-P carried the CTLA4-Ig fusion gene and CTLA4-Ig fusion gene, A-P fragment of pac gene and glu fragment of gtfB gene respectively. The expressed protein could response to specific anti-PAc antibody.


CONCLUSIONS
The targeted fusion anti-caries DNA vaccine pGJA-P is constructed successfully and expressed in eukaryotic cells correctly.
Guo
Ji-hua
JH
Key Laboratory for Oral Biomedical Engineering Ministry of Education, School of Stomatology, Wuhan University, Wuhan 430079, China.
Fan
Ming-wen
MW
Bian
Zhuan
Z
Jia
Rong
R
Chen
Zhi
Z
Peng
Bin
B
D000818:Animals; D015703:Antigens, CD; D000943:Antigens, Differentiation / Q000235:genetics / Q000276:immunology; D016466:CHO Cells; D060908:CTLA-4 Antigen; D006224:Cricetinae; D003731:Dental Caries / Q000517:prevention & control*; D005964:Glucosyltransferases / Q000235:genetics / Q000276:immunology; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D013295:Streptococcus mutans / Q000235:genetics / Q000276:immunology; D019444:Vaccines, DNA / Q000235:genetics* / Q000276:immunology
D015703:Antigens, CD; D000943:Antigens, Differentiation; D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA; D005964:Glucosyltransferases