Construction of a DNA vaccine encoding herpes simplex virus type 1 glycoprotein D
- VernacularTitle:单纯疱疹病毒I型DNA疫苗的构建
- Author:
Jianguo LI
;
Yukang YUAN
;
Jie FENG
;
Hanqing XU
;
Lin SHI
- Publication Type:Journal Article
- Keywords:
herpes simplex virus type 1;
DNA vaccine;
glycoprotein D
- From:
Journal of Xinxiang Medical College
2002;19(3):153-157
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a recombinant plasmid DNA containing herpes simplex virus type 1(HSV-1) glycoprotein D (gD) gene.Methods The HSV-1 gD gene was obtained by polymerase chain reaction (PCR) and inserted into TA cloning vector pGEM-T, then cloned into the eukaryotic expression vector pcDNA3.1 to generate pLy-D. The recombinant plasmid pLy-D, which was confirmed by partial sequencing and restriction endonuclease analysis, was transfected into Cos-7 cells and used to inoculate ICR mice via muscular injection. Immunohistochemistry and enzyme-linked immunoabsorbent assay (ELISA) were employed to test the gD expression in transfected cells and the specific anti-HSV-1 antibody in the serum of immunized mice, respectively.Results The gD eukaryotic expression plasmid pLy-D was constructed. Using the immunohistochemistry technique, the gD expression in pLy-D-transfected cells was detected. The ELISA demonstrated that specific anti-HSV-1 antibody could be induced in immunized mice after three times injection.Conclusions We constructed HSV-1 gD eukaryotic expression plasmid pLy-D which could express gD protein in transfected cells and could induce humoral immune response in mice. This observation will be helpful in designing HSV prophylactic vaccine.
