单纯疱疹病毒I型DNA疫苗的构建(英文)

Construction of a DNA vaccine encoding herpes simplex virus type 1 glycoprotein D

目的 构建表达单纯疱疹病毒 1型糖蛋白D的DNA疫苗。方法 采用聚合酶链式反应 (PCR) ,从HSV 1基因组中扩增出 gD基因 ,插入中间载体pGEM T ,然后克隆入真核表达载体pcDNA3 .1 ,构建重组质粒 pLy D .经部分测序和限制性内切酶分析证实。将pLy D转染Cos 7细胞 ,并肌肉接种ICR小鼠。用免疫组化和ELISA方法分别检测gD蛋白表达及小鼠血清中的特异性抗体。结果 转染的Cos 7细胞中有 gD蛋白的表达 ;经三次肌肉免疫接种的小鼠血清中有特异性HSV 1抗体产生。结论 本研究构建的重组质粒 pLy D能够在哺乳动物细胞中表达目的蛋白 。

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.